Spatiotemporally differential expression of HER (human epithelial growth factor receptor)-2 with sexual dimorphism in submandibular gland of mice

SUMMARY: Considering that the submandibular gland (SMG) of postnatal mice performs active cell proliferation, apoptosis and differentiation which are regulated by proto-oncogene products in cancerous cells, the expression and localization of a proto-oncogene product HER (human epidermal growth factor receptor)-2 was examined in SMG of postnatal mice. In Western blot analysis, the expression for HER-2 was high until pre-puberty, and it decreased from puberty to young adult stages with male SMG more dominant. In immunohistochemistry, the immunoreactivity was positive in acinar and ductal cells of newborn SMG with distinct localization at the intercellular apposition sites. The immunoreactivity in acinar cells progressively decreased to negligible levels by pre-pubertal stage, while it remained positive in most ductal cells throughout the postnatal time-course. The immunoreactivity in cells of terminal tubules and intercalated ducts, both of which have a high potential to produce cells, were seen at levels similar to those of more proximal ducts, while the immunoreactivity in ductal basal cells was significantly high, but the granular convoluted tubule cells were seen at negligible levels in male and at faint levels in female. In immuno-electron microscopy of excretory ducts, the immunoreactivity was dominantly localized on the basal infolding membranes as well as vesicles and vacuoles of various sizes, but rarely in Golgi apparatus and mitochondria. The immunoreactivity without association to any membranous structures were also seen, though not numerous. The relation of expression levels of HER-2 in various portions of normal SMG to those in their cancerous ones is briefly discussed.

RESUMEN: Considerando que la glándula submandibular (GSM) de ratones postnatales realiza la proliferación celular activa, apoptosis y diferenciación que están reguladas por productos protooncogénicos en células cancerosas, la expresión y localización de un producto protooncogénico HER (receptor del factor de crecimiento epidérmico humano) - 2 se examinó en GSM de estos ratones. En el análisis de Western blot, la expresión de HER-2 fue alta hasta la prepubertad, y disminuyó desde la pubertad hasta las etapas de adultos jóvenes con GSM macho más dominante. En inmunohistoquímica, la inmunorreactividad fue positiva en las células acinares y ductales de GSM de recién nacido con una localización distinta en los sitios de aposición intercelular. La inmunorreactividad en las células acinares disminuyó progresivamente a niveles insignificantes en la etapa prepuberal, mientras que permaneció positiva en la mayoría de las células ductales durante el transcurso del tiempo posnatal. La inmunorreactividad en las células de los túbulos terminales y los conductos intercalados, los cuales tienen un alto potencial para producir células, se obser- vó a niveles similares a los de los conductos más proximales, mientras que la inmunorreactividad en las células basales ductales fue significativamente alta, pero en el túbulo contorneado granular las células se observaron en niveles insignificantes en los machos y en niveles débiles en las hembras. En la microscopía inmunoelectrónica de los conductos excretores, la inmunorreactividad se localizó de manera predominante en las membranas de pliegues basales, así como en vesículas y vacuolas de varios tamaños, pero raramente en el aparato de Golgi y en las mitocondrias. También se observó la inmunorreactividad sin asociación a ninguna estructura membranosa, aunque no numerosa. Se discute brevemente la relación de los niveles de expresión de HER-2 en varias porciones de GSM normal con aquellos en sus cancerosos.

Early Postnatal Valproic Acid Exposure Increase the Protein Level of Astrocyte Markers in Frontal Cortex of Rat

OBJECTIVE: In our previous study, it has been reported that valproic acid (VPA) effects gliogenesis and increases the number of glial precursor cells during the early postnatal period. However there is no specific report that whether this process is going on up to the age of mature brain development and the consequence effect of this ongoing gliogenesis process. METHODS: As an ongoing study, using Immunoblotting analysis, we checked the level of glial protein and glial-derived factor markers in the frontal cortex of a rat brain at postnatal day (PND) 21. RESULTS: The finding of the study suggests that, in the VPA group (p < 0.05), early exposure elicited significantly to increase the expression level of glial protein cells at PND 21 in the frontal cortex of rat brain. CONCLUSION: Therefore we suggest that, alter gliogenesis and abnormal number of glial cells modulate the neurobiological dysfunction and induces the risk of neurodevelopmental disorders.

Postnatal changes in glucose transporter 3 expression in the dentate gyrus of the C57BL/6 mouse model / 한국실험동물학회지

In this study, we observed the ontogenetic changes in glucose transporter 3 (GLUT3) immunoreactivity, a major neuronal GLUT, in the dentate gyrus of mouse brains at various ages: postnatal day (P) 1, 7, 14, 28, and 56. At P1, cresyl violet staining showed abundant neurons in the dentate gyrus, whereas the granule cell layer was ill-defined. At P7, the granule cell layer was observed, and cresyl violet-positive cells were dispersed throughout the polymorphic layer. At P14, the granule cell layer was well-defined, and cresyl violet positive cells were detected abundantly in the polymorphic layer. At P28 and P56, cresyl violet-positive cells were observed in the granule cell layer, as well as in the polymorphic layer. At P1, GLUT3 immunoreactivity was detected in the dentate gyrus. At P7, GLUT3 immunoreactive cells were scattered in the polymorphic and molecular layer. However, at P14, GLUT3 immunoreactivity was observed in the polymorphic layer as well as subgranular zone of the dentate gyrus. At P28, GLUT3 immunoreactivity was detected in the polymorphic layer of the dentate gyrus. At P56, GLUT3 immunoreactivity was observed predominantly in the subgranular zone of the dentate gyrus. GLUT3 immunoreactive cells were mainly colocalized with doublecortin, which is a marker for differentiated neuroblasts, in the polymorphic layer and subgranular zone of dentate gyrus at P14 and P56. These results suggest that the expression of GLUT3 is closely associated with postnatal development of the dentate gyrus and adult neurogenesis.

Postnatal development of testis in gaddi goat (capra hircus) / Desarrollo postnatal del testículo en la cabra gaddi (capra hircus)

A study was conducted on the testicles of 30 male Gaddi goats, divided into 3 groups viz; prepubertal (0 day to 18 months), pubertal (<18 months to >5 yrs) and post- pubertal (>5 yrs) ages. The study revealed that the testis was covered by fibroserous tunica albuginia having outer fibrous layer and inner vascular layer with smooth muscle fibers at birth. The tunica albuginia continuously grew in thickness from prepubertal to postpubertal animals. The septula testis arising from the tunica albuginia, divided the parenchyma in lobules and converged at mediastinum testis. Few differentiated and undifferentiated Leydig cells were present in the intertubular spaces of neonates, which enormously increased in pubertal animals and replaced by fibroblasts in the postpubertal animals. The parenchyma comprised of solid sex cords in new born kids. These were converted into luminated tubules after 6 months of age and had clear cut tubuli contorti and tubuli recti leading to the mediastinum testis. In "0" day old kids, sex cords were filled with undifferentiated small cells located peripherally along the basement membrane and large primordial germ cells located centrally. By six months of age, latter started showing sign of degeneration and thus luminization of tubules started. Spermatogenesis startedin the seminiferous tubules of 12-18 months goat. In pubertal animals all the stages of spermatogenic cells and Sertoli cells were identified in the seminiferous tubules. The population of gonia cells and primary + secondary spermatocytes were 35% and 30% respectively. In postpubertal animals gonia cells were reduced to <30% and Primary + secondary spermatocytes were <20%. Many degenerating seminiferous tubules showed hyalinization, fibrosis and giant cells. The stroma and parenchyma ratio which was 35:65 at birth became 30:60 at 6 months, 25:75 at 12 months and 15:85 in pubertal and 20:80 in postpubertal animals. Increased stroma in last phase was associated with replacement of seminiuferous tubules by fibrous elements.

Se realizó un estudio en 30 testículos de cabras Gaddi machos divididos en 3 grupos: prepúberes (0 días a 18 meses), púberes (<18 meses y >5 años) y post-púberes (>5 años). El estudio reveló que los testículos al nacimiento estaban cubiertos por una túnica albugínea fibroserosa con una capa fibrosa externa y otra capa vascular interna con fibras musculares lisas. En los animales prepúberes hasta los pospúberes, la túnica albugínea mostró un crecimiento continuo de espesor. El septo testicular derivado de la túnica albugínea, divide el parénquima en lóbulos convergentes en el mediastino testicular. En los espacios intertubulares de los testículos de los recién nacidos, se observaron escasas células interticiales diferenciadas e indiferenciadas, sin embargo, se registró un aumento importante en los animales púberes las cuales fueron reemplazadas por fibroblastos después de la pubertad. El parénquima en los testículos de los animales recién nacidos estaba compuesto por cordones sexuales continuos. Estos se convierten en túbulos luminados después de los 6 meses de edad, evidenciando túbulos contorneados y rectos que conducen al mediastino testicular. En animales de 0 días de edad, los cordones sexuales presentaban pequeñas células no diferenciadas y situadas periféricamente a lo largo de la membrana basal y grandes células germinales primordiales céntricas. A los seis meses de edad, este último comenzó a mostrar signos de degeneración. La espermatogénesis se inició en los túbulos seminíferos desde los 12 a 18 meses de edad. En los animales púberes se identificaron en los túbulos seminíferos todas las etapas de las células de espermatogénesis y células sustentaculares. La población de células gonia fue del 35% y de espermatocitos primarios más secundarios 30%. En los animales pospúberes las células gonia se redujeron a <30% y espermatocitos primarios más secundarios a < 20%. Muchos túbulos seminíferos presentaron degeneración. La relación parénquima/estroma fue 35:65 al nacer, luego de 30:60 a los 6 meses, de 25:75 a los 12 meses, de 15:85 en animales prepúberes y 20:80 en la pubertad. El aumento del estroma en la última fase fue asociado con el reemplazo de los túbulos seminíferos por elementos fibrosos.

Myosin Va is developmentally regulated and expressed in the human cerebellum from birth to old age

Myosin Va functions as a processive, actin-based motor molecule highly enriched in the nervous system, which transports and/or tethers organelles, vesicles, and mRNA and protein translation machinery. Mutation of myosin Va leads to Griscelli disease that is associated with severe neurological deficits and a short life span. Despite playing a critical role in development, the expression of myosin Va in the central nervous system throughout the human life span has not been reported. To address this issue, the cerebellar expression of myosin Va from newborns to elderly humans was studied by immunohistochemistry using an affinity-purified anti-myosin Va antibody. Myosin Va was expressed at all ages from the 10th postnatal day to the 98th year of life, in molecular, Purkinje and granular cerebellar layers. Cerebellar myosin Va expression did not differ essentially in localization or intensity from childhood to old age, except during the postnatal developmental period. Structures resembling granules and climbing fibers in Purkinje cells were deeply stained. In dentate neurons, long processes were deeply stained by anti-myosin Va, as were punctate nuclear structures. During the first postnatal year, myosin Va was differentially expressed in the external granular layer (EGL). In the EGL, proliferating prospective granule cells were not stained by anti-myosin Va antibody. In contrast, premigratory granule cells in the EGL stained moderately. Granule cells exhibiting a migratory profile in the molecular layer were also moderately stained. In conclusion, neuronal myosin Va is developmentally regulated, and appears to be required for cerebellar function from early postnatal life to senescence.

Development of the Morphostructure and Meat Value in Chilota Lambs / Desarrollo de la Morfoestructura y el Valor Carnicero en Corderos Chilotes

Chilota sheep are typical of the Chiloé archipelago in southern Chile. The objectives of this study were: 1) To describe the development of the morphostructure of Chilota lambs in their first months of life using some variables that determine lamb meat value (zoometric traits and body weight) and 2) To determine the optimum slaughter age. Measurements were taken from 12 male single-born lambs. Body weight, Withers height; Perimeter of thorax, Chest depth, Shoulder point width and Body length were measured in each animal. The first measurements were taken at 7 days old, and the rest at 37, 67, 97, 127, 157 and 187 days old. The body weight variables and zoometric traits developed allometrically throughout the growth phase. The correlation coefficient between all the traits was very high, varying between 0.91 and 0.98. The zoometric variable that best predicted body weight was Perimeter of thorax (r2=0.93), which was fully developed in 90% of the animals at 97 days old. Maximum growth was recorded in all the studied variables during the first control month, days 7-37, but then it decreased gradually. This decrease was moderate until day 127, becoming very low from then on until end of the study. The optimum age for slaughtering fattened lambs is approximately four months, as from day 127 the daily growth rate slows down considerably, to 61.81 g/day, in comparison with the previous stage, which is 220.00 g/day. Therefore, the maintenance of lambs after four months is not productively efficient.

La oveja Chilota es una raza típica del archipiélago de Chiloé en el sur de Chile. Se midieron 12 corderos machos nacidos de parto único. En cada animal se midieronel peso corporal, la alzada a la cruz, el perímetro del tórax, diámetro dorso esternal, diámetro bicostal y la longitud corporal. Las primeras mediciones se tomaron a los 7 días de edad, y el resto a los 37, 67, 97, 127, 157 y 187 días de edad. El peso corporal y los rasgos zoométricos se desarrollaron alométricamente a lo largo de la fase de crecimiento. El coeficiente de correlaciónentre todos los rasgos fue muy elevado, variando entre 0,91 y 0,98. La variable zoométrica mejor predicha por el peso corporal fue el perímetro torácico (r2=0,93), el cual estuvo totalmente desarrollado en el 90% de los animales a los 97 días de edad. El crecimiento máximo fue registrado durante el primer mes de control (días 7-37) para todas las variables estudiadas, pero posteriormente decreció de forma gradual. Este decrecimiento fue moderado hasta el día 127, volviéndose muy lento hacia el final del estudio. La edad óptima para el sacrificio de los corderos es aproximadamente de 4 meses, dado que a partir del día 127 la tasa diaria de crecimiento disminuye considerablemente, a 61,81 g/día, en comparación con la etapa previa, donde esta tasa es de 220,00 g/día.Por lo tanto, el sacrificio de los corderos más allá de los 4 meses de edad no resulta productivamente eficiente.

Modulation of intestinal brush border membrane chemical composition during postnatal development in rats : Effect of gestational diabetes.

There was a significant increase in fucose (52%), total hexoses (16%) and hexosamine (56%) except sialic acid, which was reduced (77%) in the microvillus membrane of infants born to rat mothers made diabetic by injecting alloxan on day 3 of gestation. Expressed on the protein basis there were a significant increase in membrane, triglyceride, total cholesterol, and phospholipids content of brush border in pups from diabetic group between 5-45 days of postnatal age. Intestinal morphology in diabetic group showed, regression of tubular glands, distorted cellular organization of mucosal cells, reduction in the mucosal cell height and number of secretory goblet cells. These findings suggest that the gestational diabetes affects the sugar and lipid composition of the intestinal brush border membrane in rats during early stages of the postnatal development, which may be associated with compromised tissue functions later in life.

Histochemical study of lectin-binding patterns in the rat vomeronasal organ during postnatal development / 대한수의학회지

Histochemical patterns of lectin binding during development of the rat vomeronasal organ (VNO) were studied to determine whether glycoconjugates are differently expressed after birth. Three types of lectins, Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA-I), were studied histochemically in the rat VNO at various stages post-birth: postnatal days 1 and 7, the preweaning period (4 weeks after birth), and at sexual maturity (8 weeks after birth). The free border of the vomeronasal sensory epithelium was positive for both WGA and UEA-I in rats of all ages; whereas, VNO receptor cells and supporting cells were positive only for both WGA and UEA-I from 4 weeks after birth. DBA reactivity was detected in the free border but less so in receptor cells and supporting cells. WGA and UEA-I, but not DBA, showed similar patterns in various ages. In the Jacobson's gland, WGA, UEA-I and DBA were detected in some acini from 4 weeks after birth but not at postnatal days 1 or 7. Collectively, reactivity for three lectins, WGA, UEA-I and DBA, increased in receptor cells and gland acini during postnatal development, possibly contributing to the enhanced chemoreception in rats.

Detailed Differentiation of Calbindin D-28k-Immunoreactive Cells in the Dentate Gyrus in C57BL/6 Mice at Early Postnatal Stages / 한국실험동물학회지

The hippocampus makes new memories and is involved in mental cognition, and the hippocampal dentate gyrus (DG) is critical because neurogenesis, which occurs throughout life, occurs in the DG. We observed the differentiation of neuroblasts into mature neurons (granule cells) in the DG of C57BL/6 mice at various early postnatal (P) ages: P1, P7, P14, and P21 using doublecortin (DCX) immunohistochemistry (IHC) for neuroblasts and calbindin D-28k (CB) IHC for granule cells. DCX-positive cells decreased in the DG with age; however, CB+ cells increased over time. At P1, DCX and CB double-labeled (DCX+CB+) cells were scattered throughout the DG. At P7, DCX+CB+ cells (about 92% of CB+ cells) were seen only in the granule cell layer (GCL) of the dorsal blade. At P14, DCX+CB+ cells (about 66% of CB+ cells) were found in the lower half of the GCL of both blades. In contrast, at P21, about 18% of CB+ cells were DCX+CB+ cells, and they were mainly located only in the subgranular zone of the DG. These results suggest that the developmental pattern of DCX+CB+ cells changes with time in the early postnatal stages.

Differential patterns of nestin and glial fibrillary acidic protein expression in mouse hippocampus during postnatal development

Intermediate filaments, including nestin and glial fibrillary acidic protein (GFAP), are important for the brain to accommodate neural activities and changes during development. The present study examined the temporal changes of nestin and GFAP protein levels in the postnatal development of the mouse hippocampus. Mouse hippocampi were sampled on postnatal day (PND) 1, 3, 6, 18, and 48. Western blot analysis showed that nestin expression was high at PND 1 and markedly decreased until PND 18. Conversely, GFAP expression was acutely increased in the early phase of postnatal development. Nestin immunoreactivity was localized mainly in the processes of ramified cells at PND 1, but expression subsequently decreased. In contrast, GFAP was evident mainly in the marginal cells of the hippocampus at PND 1, but immunoreactivity revealed satellite, radial, or ramified shapes of the cells from PND 6-48. This study demonstrates that the opposing pattern of nestin and GFAP expressions in mouse hippocampus during postnatal development occur in the early development stage (PND 1-18), suggesting that the opposing change of nestin and GFAP in early postnatal development is important for neural differentiation and positioning in the mouse hippocampus.

Study of sensory-motor and somatic development of the offspring of rats (wistar) treated with caffeine / Estudo da sensório-motora e desenvolvimento somático da prole de ratos (wistar) tratados com cafeína

The influence of caffeine, administered to rats, on the somatic and sensory-motor development of the offspring was investigated. Female Wistar rats were divided into a control group and a treated group and received drinking water and a 0.1 percent solution of caffeine orally, respectively. The offspring, also divided into a control group and a treated group, received daily monitoring until the 20th day of life to verify alterations in somatic neural development. The offspring of the treated group had reduced weight on the day of birth and on the 1st, 5th, 15th and 20th days of life; shorter snout-anus length (evaluation done daily); shorter snout-tail length on the day of birth and on the 1st, 5th and 10th days of life, and signs of retardation of somatic and sensory-motor maturation. These results allowed the conclusion that administration of caffeine to rats affects somatic and sensory-motor development of offspring.

Estudou-se a influência da cafeína, administrada a ratos, no desenvolvimento somático e sensorial-motor da prole. Ratos Wistar fêmeas foram divididos em grupo controle e grupo tratado e receberam água e solução de cafeína a 0,1 por cento, respectivamente. A prole, também dividida em grupo controle e grupo tratado, foi monitorada diariamente até o 20º. dia de vida para se observar as alterações no desenvolvimento somático neural. O grupo tratado apresentou peso reduzido no dia do nascimento e nos 1º, 5º., 15º e 20º. dias de vida; comprimento focinho-ânus mais curto (avaliação efetuada diariamente); comprimento focinho-cauda mais curto no dia do nascimento e nos 1º, 5º., 15º e 20º. dias de vida e sinais de retardamento da maturação somática e sensorial-motora. Esses resultados permitem que se conclua que a administração da cafeína a ratos afeta o desenvolvimento somático e sensorial-motor da prole.

The expression and localization of inhibin isotypes in mouse testis during postnatal development

Inhibin, which is important for normal gonadal function, acts on the pituitary gonadotropins to suppress folliclestimulating hormone (FSH) secretion. The level and cellular localization of the inhibin isotypes, alpha, beta(A) and beta(B), in the testis of mice were examined during postnatal development in order to determine if inhibin expression is related to testicular maturation. Mouse testes were sampled on postnatal days (PNDs) 1, 3, 6, 18, 48 and 120, and analyzed by Western blotting and immunofluorescence. Western blot analysis showed very low levels of inhibin alpha, beta(A) and beta(B) expression in the testes at days 1 to 6 after birth. The levels then increased gradually from PND 18 to 48-120, and there were significant peaks at PND 48. Inhibin alpha, beta(A) and beta(B) were detected in testicular cells during postnatal development using immunohistochemistry. The immunoreactivity of inhibin alpha was rarely observed in testicular cells during PND 1 to 6, or in the cytoplasmic process of Sertoli cells surrounding the germ cells and interstitial cells during PND 18 to 120. Inhibin beta(A) and beta(B) immunoreactivity was rarely observed in the testis from PND 1 to 6. On the other hand, it was observed in some spermatogonial cells, as well as in the interstitial space between PND 48 and PND 120. We conclude that the expression of inhibin isotypes increases progressively in the testis of mice with increasing postnatal age, suggesting that inhibin is associated with a negative feedback signal for FSH in testicular maturation.

Effect of Maternal Thyroxine Treatment on the Postnatal Development of Brain-derived Neurotrophic Factor-containing Neuron in the Brain of Pups of Alcohol Abused Mother / 대한해부학회지

Maternal alcohol abuse is thought to be the common cause of mental retardation. Especially, continuous alcohol consumption during critical period of brain development induce fetal alcohol effects. In this study, the authors investigated the effects of maternal alcohol drinking on the postnatal changes of BDNF contents and patterns of BDNF-containing neuron in neonatal rat brain, and, the influence of maternal thyroxine treatment on the brain of pups of alcohol abused mother. Pregnant rats were divided into three groups. Alcohol-fed group (n=4) received 35 calories of liquid alcohol diet daily from gestation day 6; control pair-fed group (n=4) was fed a liquid diet in dextrin replaced alcohol isocalorically; alcohol+T4 group (n=4) received 35 calories liquid alcohol diet and exogenous thyroxine (5 microgram/kg/day) subcutaneously. The amount of BDNF was significantly higher in the alcohol+T4 group as compared to the alcohol group at P7, P14 and P21, especially, alcohol+T4-exposed pups showed a significant increase of BDNF at P7. The decrease in BDNF was found in alcohol group compared to control pair-fed group at all ages. In alcohol+T4 group, BDNF-containing Purkinje cells exhibited mature pattern and monolayer arrangement at P14. Alcohol+T4 group showed mature pattern and numerical increase of BDNF-containing cells in cerebral cortex, hypothalamus and hippocampus at P7. The BDNF immunoreactivity of hippocampus continued to show prominent configuration in alcohol+T4 group at P28. These results indicate that the increase of the BDNF-containing neurons and BDNF amount in pups of thyroxinesupplemented alcohol-exposed dams as compared to control pair-fed and alcohol-exposed pups at P7, presumably suggest the early postnatal growth stimulatory effect of the exogenously supplemented thyroxine. Therefore, the increase of BDNF synthesis caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects, one of the ill effects as a result of the dysthyroid state following maternal alcohol abuse.

Effect of Exogenous Thyroxine on the Postnatal Development of Serotonin-containing Neuron in Fetal Alcohol Effects in the Rat Raphe Nuclei / 체질인류학회지

Serotonin (5-hydroxytryptamine, 5-HT) has been concerned in the pathophysiology of various neuropsychiatric disorders. It is known to modulate emotion, cognition, endocrine activity, motor function, and pain. In the present study, the effects of exogenous thyroxine (T4) on the postnatal development of serotonin-containing neuron in the rat raphe nuclei with fetal alcohol effects were investigated using immunohistochemistry. These experimental animals were divided into three groups : the alcohol-fed group received 35 calories liquid ethanol diet; the control pair-fed group was fed a liquid diet in dextrin replaced alcohol isocalorically; alcohol+T4 group received alcohol diet and exogenous thyroxine subcutaneously. After the pups were born, the pups of each were fostered by surragate mother. An average of four pups, one from each litter, were killed at days 0, 7, 14, 21, and 28 for each of the above three groups. As a result, in alcohol group, serotonin-immunoreactivity was weakly stained at all postnatal ages compared to control pair-fed and alcohol+T4 group. The intensity of serotonin immunoreactivity was more prominent in alcohlol+T4 group than in control pair-fed group at P0. Mature patterns of serotonin-containing neurons were observed in control pair-fed and alcohol+T4 group at P7. A similar developmental pattern of serotonin-containing neuron was observed on and after P7 in control pair-fed and alcohol+T4 group. These results suggest that the increase of serotonin synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects, one of the ill effects as a result of the dysthyroid state following maternal alcohol abuse.

Postnatal Developmental Expression of TEM7 mRNA and its Putative Ligand in the Rat Cerebellum / 대한해부학회지

Tumor endothelial marker 7 (TEM7) is a putative transmembrane protein that is highly expressed in the tumor endothelium and cerebellar neurons. In the present study, the expression profile of TEM7 mRNA and its putative ligand in the developing cerebellum of the rat was investigated using in situ hybridization and ligand binding assay. The secreted recombinant ectodomain of TEM7 was employed to label the expression of putative ligand of TEM7 in the cerebellum. The expression of a putative ligand of TEM7 demonstrated by using TEM7 ectodomain was found in the molecular layer of the cerebellum, where the dendritic trees of Purkinje cells are present. A developmental study has shown that TEM7 mRNA expression in the Purkinje neurons was increased with age during postnatal development, whereas the putative ligand labeling in the molecular layer was observed throughout the developmental period. These findings indicate that TEM7-ligand interaction plays a role in the differentiation of Purkinje cells during postnatal development.

Effect of Maternal Thyroxine Treatment on the Postnatal Development of Brain-derived Neurotrophic Factor-containing Purkinje Cell in Fetal Alcohol Effects in the Rat Cerebellum / 체질인류학회지

Maternal alcohol abuse is thought to be the common cause of mental retardation. Even moderate maternal alcohol consumption may produce fetal alcohol effects with behavioral and learning difficulties, if the drinking is associated with malnutrition. Especially, continuous alcohol consumption during critical period of brain development is very likely to produce fetal alcohol effects. The aims of this study are to investigate whether exogenous thyroxine treatment to alcohol -fed dams may ameliorate the detrimental effects of alcohol on the postnatal development of BDNF -containing Purkinje cell of the cerebellar cortex of the offspring. The morphological features of the growth and maturation were observed at 0, 7, 14, 21, 28 postnatal days via immunohistochemistry. In addition, electron microscopic finding of BDNF -containing Purkinje cell at P14 was also examined. Time -pregnant rats were divided into three groups. Alcohol -fed group received 35 calories of liquid alcohol diet daily from gestation day 6; control pair -fed group was fed a liquid diet in which dextrin replaced alcohol isocalorically; alcohol +/-T4 group received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. As a result, a similar developmental pattern of BDNF -immunoreactive Purkinje cells was observed in control pair - fed and alcohol+/-T4 group on and after P14. These cells of alcohol -fed group showed immature features. Single -layer arrangement of these cells in alcohol -fed group was not completely achieved throughout postnatal life. Electron microscopic observations of BDNF -immunoreactive Purkinje cells at P14 revealed large nucleus, small cytoplasm, small amount of ribosomal collection and rudimentary cytoplasmic organelles in alcohol -fed group. The morphology of BDNF -immunoreactive Purkinje cell in alcohol +/-T4 group was similar to that in control pair -fed group. It was characterized by numerous short segments of rough endoplasmic reticulum, many of which showed a tendency of parallel alignment that suggested an attempt at Nissl body configuration. The cytology of Golgi complexes was also found within the cytoplasm in perinuclear location. Those observed differences of postnatal maturation patterns between alcohol -fed and alcohol +/-T4 group may indicate the beneficial effects on the postnatal development of BDNF -containing Purkinje cells in cerebellar cortex in the pups of thyroxine -treated alcohol -exposed dams. These results suggest that the increase of BDNF synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects as a result of the dysthyroid state following maternal alcohol abuse.

Roles of Stem Cell Factor and c-kit Receptor in the Development of Cerebellar GABAergic Neurons / 대한해부학회지

Evidence that Stem cell factor (SCF) and c-Kit receptor tyrosine kinase are expressed in the cerebellum during postnatal development, suggests a possible contribution of the SCF/Kit signaling pathway in the cerebellar development. In the present study, we prepared cerebellar cultures from C57Bl/6J mouse at postnatal day 1and 7 to investigate the role of c-kit receptor and SCF in regulation of growth and differentiation in the postnatal cerebellar GABAergic cells. SCF increased the number of survival cerebellar cells and density of glutamic acid decarboxylase 65/67 (GAD65/67) and calbindin D-28K expression in the immunoblot analysis. SCF also improved the neurite extension of the interneuron neuritis and dendritogenesis of Purkinje cells. Treatment with c-Kit antibody accelerated cellular loss in serum-free media and decreased the growth ability and dendritogenesis of Purkinje cells and cerebellar inhibitory interneurons. Our data suggest that SCF and c-kit receptor are required for the normal growth of postnatal cerebellum and a possible involvement of functional regulation through the SCF/c-kit receptor pathways in the postnatal cerebellar development.

Postnatal Development of Brain Natriuretic Peptide-immunoreactive Neuron in the Hypothalamus of the Rat / 체질인류학회지

Brain natriuretic peptide (BNP) is a neuropeptide, isolated from porcine brain that is homologous with atriopeptin. Magnocellular neurosecretory cells located in the paraventricular nucleus and supraoptic nucleus synthesize and secrete neurohormones. The purpose of this study was to investigate distribution of BNP immunoreactivity throughout the rat hypothalamus from the day of birth to 30 days and adult using immunoperoxidase and immunofluorescent staining. The first BNP immunoreactive neurons appeared in the paraventricular and supraoptic nucleus at P10. In adult, BNP immunoreactivity was widely distributed throughout regions of the hypothalamus including dorsomedial hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus and internal layer of median eminence. The intensity of BNP immunoreactivity was weak in almost all hypothalamic nuclei except the paraventricular and supraoptic nuclei. BNP immunoreactivity was first observed in the lateral hypothalamic area at P15. In retrochiasmatic supraoptic nucleus, BNP immunoreactivity was first observed at P20 and remarkably distributed in adult. In the present study, distinct localization of BNP immunoreactivity was in the hypothalamic cell bodies and fibers. Although the role of BNP in the brain is yet to be determined, these results indicate that BNP in the neurons of hypothalamus play important role in the regulation of a variety of neurosecretory functions as a neuromodulator during postnatal development of the hypothalamus.

Postnatal Development of Brain Natriuretic Peptide-immunoreactive Neuron in the Hypothalamus of the Rat / 체질인류학회지

Brain natriuretic peptide (BNP) is a neuropeptide, isolated from porcine brain that is homologous with atriopeptin. Magnocellular neurosecretory cells located in the paraventricular nucleus and supraoptic nucleus synthesize and secrete neurohormones. The purpose of this study was to investigate distribution of BNP immunoreactivity throughout the rat hypothalamus from the day of birth to 30 days and adult using immunoperoxidase and immunofluorescent staining. The first BNP immunoreactive neurons appeared in the paraventricular and supraoptic nucleus at P10. In adult, BNP immunoreactivity was widely distributed throughout regions of the hypothalamus including dorsomedial hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus and internal layer of median eminence. The intensity of BNP immunoreactivity was weak in almost all hypothalamic nuclei except the paraventricular and supraoptic nuclei. BNP immunoreactivity was first observed in the lateral hypothalamic area at P15. In retrochiasmatic supraoptic nucleus, BNP immunoreactivity was first observed at P20 and remarkably distributed in adult. In the present study, distinct localization of BNP immunoreactivity was in the hypothalamic cell bodies and fibers. Although the role of BNP in the brain is yet to be determined, these results indicate that BNP in the neurons of hypothalamus play important role in the regulation of a variety of neurosecretory functions as a neuromodulator during postnatal development of the hypothalamus.

Immunohistochemical Study on Platelet-Derived Growth Factor alpha-Receptor (PDGF -alpha R) in Developing Canine CNS / 대한해부학회지

Platelet-derived growth factor (PDGF) was initially described for its mitogenic activity on smooth muscle cells, fibroblast, and glial cells. The biological activities of PDGF include stimulation of mitogenesis, differentiation, wound healing, inflammation, and tumor formation. The localization of platelet-derived growth factor-alpha Receptor (PDGF-alpha R) in central nervous system was commonly restricted to oligodendrocyte progenitors during late embryonic and postnatal development. However, several studies recently demonstrated that postnatal neurons could also synthesize PDGF-alpha R in rodents. In the present study, to analyze the distributional pattern of PDGF-alpha R during postnatal development of the canine CNS, we used immunohistochemical method on sections of canine brain tissue. We found that neurons of various CNS regions, including cerebral cortex, striatum, diencephalon, nuclei of brain stem, cerebellum, spinal cord, exhibited the immunoreactivity to PDGF-alpha R as early as postnatal day 0. Generally PDGF-alpha R immunoreactivity was well localized in the dendrites and axons of neuron during the postnatal day 14 and postnatal day 28, and then showed diminished pattern. But neuronal immunoreactivity to PDGF-alpha R were maintained postnatal 6 month. These results suggest that the localization of PDGF-alpha R in postnatal developing neurons supports the several roles of PDGF for neurons including maturation and survival.