Melatonin is involved in the modulation of the hypothalamic and pituitary activity in the South American plains vizcacha, Lagostomus maximus.

Melatonin, the key messenger of photoperiodic information, is synthesized in the pineal gland by arylalkylamine N-acetyltransferase enzyme (AANAT). It binds to specific receptors MT1 and MT2 located in the hypothalamus and pituitary gland. Melatonin can modulate the reproductive axis affecting the secretion of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH). The South American plains vizcacha, Lagostomus maximus, shows natural poliovulation of up to 800 oocytes per estrous cycle, a 154-day long pregnancy, and reactivation of the reproductive axis at mid-gestation with pre-ovulatory follicular recruitment, presence of active corpora lutea, and variations of the endocrine status. Here we analyzed the involvement of melatonin in the modulation of the hypothalamic and pituitary gland physiology of vizcacha thorough several approaches, including histological localization of melatoninergic system components, assessment of melatoninergic components expression throughout the reproductive cycle, and evaluation of the effect of melatonin on hypothalamic and pituitary activities during the follicular and luteal phases of the estrous cycle. AANAT and melatonin receptors were localized in the pineal gland and preoptic area of the hypothalamus. Increase in pineal AANAT and serum melatonin expression was observed as pregnancy progressed, with the lowest hypothalamic MT1 and MT2 levels at mid-pregnancy. Pulsatility assays demonstrated that melatonin induces GnRH and LH secretion at luteal phase. The melatoninergic system effects on hypothalamic and pituitary gland hormones secretion during pregnancy pinpoint to melatonin as a potential key factor underlying the reactivation of the reproductive axis activity at mid-gestation.

First record of an infection by tissue cyst-forming coccidia in wild vizcachas (Lagostomus maximus, Rodentia) of Argentina.

Endoparasites of the Sarcocystidae family share the ability to form tissue cysts in their intermediate hosts, ultimately leading to pathogenesis in the definitive hosts that include various mammals, reptiles and birds. In our research on the endocrinology of the female vizcachas (Lagostomus maximus), we have found abnormal cystic structures in the ovaries of some individuals. So far, no cases of infection by tissue cyst-forming parasites have been reported in this species. To evaluate whether this autochthonous wild rodent is an intermediate host of an undescribed endoparasite, histological sections from various organs were examined. Pinhead-sized tissue cysts were found in the ovaries, mammary glands, uterus, pituitary, brain, adrenals and spleen, of both pregnant and non-pregnant females. The presence of cysts in the adult brain and embryonic tissue is indicative of the ability of the parasite to cross both the blood-brain and placental barriers. The infected brains exhibited a lower cyst density than that seen in other organs. Regardless of their location in superficial or deep tissue, the cysts were surrounded by a layer of connective tissue. Histologically, the cyst wall consisted of an outer layer of fibroblasts and collagen fibers, and an inner, granular-looking layer composed of host nucleated cells surrounding thousands of spindle-shaped bradyzoites. Outside the cysts, the host cellular structures showed normal appearance. The remarkable morphological similarities between the cysts studied here with those reported in naturally infected rabbits from an area neighboring the one inhabited by the vizcachas point to Besnoitia sp. as a plausible candidate. More studies will be necessary to confirm the identity of the parasite. Nevertheless, this is the first report of L. maximus as an intermediate host for a tissue cyst-forming coccidia.

Pituitary estrogen receptor alpha is involved in luteinizing hormone pulsatility at mid-gestation in the South American plains vizcacha, Lagostomus maximus (Rodentia, Caviomorpha).

The South American plains vizcacha, Lagostomus maximus, is a caviomorph rodent native from Argentina, Bolivia and Paraguay. It shows peculiar reproductive features like pre-ovulatory follicle recruitment during pregnancy with an ovulatory process at around mid-gestation. We have described the activation of the hypothalamic - pituitary - ovarian (HPO) axis during pregnancy. A progressive decrease of progesterone (P4) at mid-pregnancy elicits the delivery of gonadotropin-releasing hormone (GnRH) with the consequent secretion of follicle stimulating hormone (FSH) and estradiol (E2) followed by luteinizing hormone (LH) release resulting in follicular luteinization and the P4 concentration recover. Pituitary gland is the central regulator of the HPO axis being E2 a key hormone involved in the regulation of its activity. In this work we analyzed the action of E2 on the pituitary response to the GnRH wave as well as its involvement on LH secretion at mid-gestation in L. maximus. The expression of GnRHR at the pituitary pars distalis showed a significant decrease at mid-pregnancy compared to early- and term-gestating females. ERα showed a significant increment from mid-gestation whereas ERß did not show variations throughout pregnancy; whereas the LH expression in the pituitary pars distalis showed a significant increase at mid-gestation, concordantly with serum LH, which was followed by a decrease at term-gestation with similar values than at early-pregnancy. The number of cells with co-localization of ERα and GnRHR showed a decline at mid-pregnancy related to early- and term-gestation, whereas the cells with co-localization of ERα and LH increased at mid- and term-pregnancy. On the other hand, ex vivo measuring of LH pulsatility showed a significant increment in the total mass of LH delivered at mid-pregnancy followed by a decrease at term-gestation. The stimulation of ERα with the PPT specific agonist induced a significant increment in the total mass of LH released, whereas no changes were determined when ERß was stimulated with its specific agonist MPP. These results suggest that LH pulsatility rise at mid-pregnancy would be enabled by the increase of E2 acting through ERα.

Local production of neurostradiol affects gonadotropin-releasing hormone (GnRH) secretion at mid-gestation in Lagostomus maximus (Rodentia, Caviomorpha).

Females of the South American plains vizcacha, Lagostomus maximus, show peculiar reproductive features such as massive polyovulation up to 800 oocytes per estrous cycle and an ovulatory process around mid-gestation arising from the reactivation of the hypothalamic-hypophyseal-ovary (H.H.O.) axis. Estradiol (E2) regulates gonadotropin-releasing hormone (GnRH) expression. Biosynthesis of estrogens results from the aromatization of androgens by aromatase, which mainly occurs in the gonads, but has also been described in the hypothalamus. The recently described correlation between GnRH and ERα expression patterns in the hypothalamus of the vizcacha during pregnancy, with coexpression in the same neurons of the medial preoptic area, suggests that hypothalamic synthesis of E2 may affect GnRH neurons and contribute with systemic E2 to modulate GnRH delivery during the gestation. To elucidate this hypothesis, hypothalamic expression and the action of aromatase on GnRH release were evaluated in female vizcachas throughout pregnancy. Aromatase and GnRH expression was increased significantly in mid-pregnant and term-pregnant vizcachas compared to early-pregnant and nonpregnant females. In addition, aromatase and GnRH were colocalized in neurons of the medial preoptic area of the hypothalamus throughout gestation. The blockage of the negative feedback of E2 induced by the inhibition of aromatase resulted in a significant increment of GnRH-secreted mass by hypothalamic explants. E2 produced in the same neurons as GnRH may drive intracellular E2 to higher levels than those obtained from systemic circulation alone. This may trigger for a prompt GnRH availability enabling H.H.O. activity at mid-gestation with ovulation and formation of accessory corpora lutea with steroidogenic activity that produce the necessary progesterone to maintain gestation to term and guarantee the reproductive success.

Comparative Analysis of the Morphology, Ultrastructure, and Glycosylation Pattern of the Jejunum and Ileum of the Wild Rodent Lagostomus maximus.

Morphological and histochemical analyses were performed to characterize the histology, ultrastructure, and glycosylation pattern of the jejunum and ileum of the wild rodent Lagostomus maximus. Enterocytes, goblet cells, Paneth cells, and enteroendocrine cells were identified in both intestinal epithelia. Two morphological types of enterocytes were identified only in the ileum based on their cytoplasm electron density. Although the histological and ultrastructural examination showed that the epithelia of both anatomical regions were morphologically similar, a certain specialization in their secretory products was evident. The glycosylation pattern of the jejunum and ileum was characterized in situ by histochemical and lectin histochemical methods. Histochemical results revealed the presence of carboxylated and sulfated gycoconjugates in both regions, although sulfomucins were clearly prevalent in the ileum. Sialic acid was highly O-acetylated and particularly abundant in the jejunum. The KOH/PA*/Bh/PAS technique evidenced a more intense histochemical reaction in the jejunal than in the ileum goblet cells, demonstrating a reduction of neutral mucin secretion in the distal small intestine. Further specific differences were revealed by lectin histochemistry. These data evidenced that the nature of mucus varies at different anatomical regions, probably adapted to physiological requirements.

Variation in progesterone receptors and GnRH expression in the hypothalamus of the pregnant South American plains vizcacha, Lagostomus maximus (Mammalia, Rodentia).

In mammals, elevated levels of progesterone (P4) throughout gestation maintain a negative feedback over the hypothalamic-hypophyseal-gonadal (H-H-G) axis, avoiding preovulatory follicular growth and preventing ovulation. Recent studies showed that in the South American plains vizcacha (Lagostomus maximus) folliculogenesis progresses to preovulatory stages during gestation, and an ovulatory process seems to occur at midgestation. The aim of this work was to analyze hypothalamic gonadotropin-releasing hormone (GnRH) and P4 receptors (PR) expression and luteinizing hormone (LH) secretion and correlate these with the functional state of the ovary in nonovulating and ovulating females and gestating females with special emphasis in the supposedly ovulating females at midgestation. We investigated P4 and LH serum levels as well as the distribution, localization, and expression of PR and GnRH in the hypothalamus of L. maximus at different time points during gestation and in nongestating, ovulating and nonovulating, females. A significant increment in GnRH, P4, and LH was detected in midpregnant vizcachas with respect to early-pregnant and to ovulating females. PR was also significantly increased in midpregnant animals. PR was detected in neurons of the preoptic and hypothalamic areas. Coexistence of both PR and GnRH in neurons of medial preoptic area and supraoptic nucleus was detected. Midpregnant animals showed increased number of PR immunoreactive cells at median eminence, localized adjacently to GnRH immunoreactive fibers. High expression of hypothalamic GnRH and PR, despite an increased level of P4, was correlated with the presence of antral, preovulatory follicles, and luteinized unruptured follicles at midgestation that suggest a possible role of the H-H-G axis in the modulation of ovulation during gestation in L. maximus.

Forma e função dos músculos perineais da viscacha (Lagostomus maximus) / Shape and function of the perineal muscles of viscacha (Lagostomus maximus)

Entre as espécies de roedores já estudadas podemos destacar a grande variação na morfologia do aparelho reprodutor masculino. Assim, considerando a importância ecológica dos roedores, e a grande representatividade numérica quanto geográfica destes animais, bem como a escassez que aborda a anatomia reprodutiva, desenvolvemos este trabalho com um roedor histricomorfo da América do Sul, a viscacha. Este animal apresenta algumas características reprodutivas bem peculiares, desta forma descreveremos a anatomia macroscópica da musculatura perineal, e o papel dos músculos no comportamento copulatório desta espécie. A região perineal da viscacha é composta por cinco músculos, sendo que três músculos encontram-se dispostos no diafragama urogenital superficial, Musculus ischiocavernosus, M. bulbocavernosus e M. bulbospongiosus, e pelos músculos que se encontram no diafragma pélvico, M. levator ani e M. retractor penis. Sendo assim, destacamos que o estudo do assoalho pélvico em animais silvestres é de grande valia, uma vez que contribuem com o melhor entendimento dos mecanismos relacionados â ereção e ejaculação; ou seja colaboram com os estudos sobre a reprodução dos animais.

Among the rodent species studied we can highlight the wide variation in the morphology of the male reproductive system. Thus, considering the ecological importance of rodents, and the large number and geographical representation of this animal, as well as shortages regarding the reproductive anatomy, we developed this study with viscacha, a South American histricomorph rodent. As this species has some very peculiar reproductive features, we described the gross anatomy of the perineal muscles and the role of copulatory behavior. The perineal region of viscacha is composed of five muscles, three of which are arranged in the superficial genitourinary diaphragm, as Musculus ischiocavernosus, M. bulbocavernosus and M. bulbospongiosus, and the muscles that lie at the pelvic diaphragm, M. levator ani and M. retractor penis. Therefore, we emphasize that the study of the pelvic floor in wild animals is of great value, then contribute to a better understanding of the mechanisms related to erection and ejaculation or collaborate with studies on the reproduction of animals.

Forma e função dos músculos perineais da viscacha (Lagostomus maximus) / Shape and function of the perineal muscles of viscacha (Lagostomus maximus)

Entre as espécies de roedores já estudadas podemos destacar a grande variação na morfologia do aparelho reprodutor masculino. Assim, considerando a importância ecológica dos roedores, e a grande representatividade numérica quanto geográfica destes animais, bem como a escassez que aborda a anatomia reprodutiva, desenvolvemos este trabalho com um roedor histricomorfo da América do Sul, a viscacha. Este animal apresenta algumas características reprodutivas bem peculiares, desta forma descreveremos a anatomia macroscópica da musculatura perineal, e o papel dos músculos no comportamento copulatório desta espécie. A região perineal da viscacha é composta por cinco músculos, sendo que três músculos encontram-se dispostos no diafragama urogenital superficial, Musculus ischiocavernosus, M. bulbocavernosus e M. bulbospongiosus, e pelos músculos que se encontram no diafragma pélvico, M. levator ani e M. retractor penis. Sendo assim, destacamos que o estudo do assoalho pélvico em animais silvestres é de grande valia, uma vez que contribuem com o melhor entendimento dos mecanismos relacionados â ereção e ejaculação; ou seja colaboram com os estudos sobre a reprodução dos animais.(AU)

Among the rodent species studied we can highlight the wide variation in the morphology of the male reproductive system. Thus, considering the ecological importance of rodents, and the large number and geographical representation of this animal, as well as shortages regarding the reproductive anatomy, we developed this study with viscacha, a South American histricomorph rodent. As this species has some very peculiar reproductive features, we described the gross anatomy of the perineal muscles and the role of copulatory behavior. The perineal region of viscacha is composed of five muscles, three of which are arranged in the superficial genitourinary diaphragm, as Musculus ischiocavernosus, M. bulbocavernosus and M. bulbospongiosus, and the muscles that lie at the pelvic diaphragm, M. levator ani and M. retractor penis. Therefore, we emphasize that the study of the pelvic floor in wild animals is of great value, then contribute to a better understanding of the mechanisms related to erection and ejaculation or collaborate with studies on the reproduction of animals.(AU)

Inmovilización química y evaluación de salud de vizcachas salvajes (Lagostomus maximus) en el chaco arido argentino / Chemical restraint and sanitary assessment of wild vizcachas (Lagostomus maximus) in the arid argentin chaco / Contenção farmacológica e avaliação sanitária de Viscachas selvagens (Lagostomus maximus) no chaco árido argentino

Con el objetivo de evaluar la salud de vizcachas salvajes (Lagostomus maximus), se capturaron en el Chaco Árido argentino 10 ejemplares y se efectuaron 11 inmovilizaciones con anestesias fijas o inhalatoria. Para las anestesias, se administraron una combinación de tiletamina- zolazepam (n = 7) o una combinación de medetomidina y ketamina (n = 2) o gas isofluorano con un vaporizador portátil creando un circuito semicerrado (n = 2). Los estudios de salud comprendieron hematología, bioquímica sanguínea, análisis parasitológicos y de exposición a agentes infecciosos mediante cultivo y serología. El uso de tiletamina-zolazepam resultó en una pobre calidad anestésica y recuperaciones prolongadas. Por El contrario, el uso de medetomidina-ketamina o de isofluorano, proporcionaron inmovilizaciones más seguras. Se concluye que el protocolo testado de tiletamina-zolazepam no sería recomendable para esta especie, sobre todo en vida libre. La búsqueda de anticuerpos para los agentes infecciosos evaluados resultó negativa, por otra parte, se aisló Listeria innocua serovar 6a em hisopados rectales, se constató infestación con pulgas (Pulex irritans), ooquistes de Eimeria lagostomi, huevos de la Familia Trichostrongylidae y huevos similares a Heteroxynema viscaciae(AU)

Com objetivo de avaliar a condição sanitária de viscachas selvagens (Lagostomus maximus), foram capturados, no Chaco Árido argentino, 10 animais e realizadas 11 imobilizações, com anestesia injetável ou inalatória. Empregaram-se três diferentes protocolos: tiletamina-zolazepam (n = 7), metetomidina-cetamina (n = 2) ou isoflurano com vaporizador portátil em sistema semi-fechado (n = 2). A avaliação sanitária consistiu de hematologia, bioquímica sérica, exames parasitológicos e avaliação da exposição a agentes infecciosos por cultura e sorologia. O uso de tiletamina-zolazepam proporcionou má qualidade anestésica e recuperações prolongadas, enquanto a combinação medetomidina-cetamina, possibilitou imobilizações mais seguras. Assim, o protocolo testado de tiletamina-zolazepam não é indicado para esta espécie, especialmente em animais de vida livre. A pesquisa de anticorpos para os agentes infecciosos avaliados foi negativa, mas isolou-se Listeria innocua serovar 6a de suabes retais, e foram encontradas pulgas (Pulex irritans), oocistos de Eimeria lagostomi, ovos de Trichostrongylidae e ovos similares a Heteroxynema viscaciae.(AU)

In order to evaluate the sanitary condition of wild vizcachas (Lagostomus maximus), 10 animals were captured in the Arid Chaco of Argentina and 11 immobilizations were performed through injectable or volatile anesthesia. Three different protocols were used: tiletamine-zolazepam (n = 7), medetomidine-ketamine (n = 2), or isofluorane with a portable vaporizer in a semi-closed circuit (n = 2). The sanitary assessment consisted of hematology, blood chemistry, parasitological analysis, and the assessment of the exposure to infectious agents by culture and serology. The use of tiletamine-zolazepam caused poor anesthetic quality and prolonged recoveries, while the medetomidine-ketamin combination, however, caused safer immobilizations. Thus, the use of the tested protocol of tiletamine-zolazepam is not advisable for this species, mainly for free-ranging animals. The search of antibodies for the evaluated infectious agents was negative, but there was isolated Listeria innocua serovar 6a from rectal swabs , fleas were found (Pulex irritans), oocytes of Eimeria lagostomi, Trichostrongylidae eggs as well as similar Heteroxynema viscaciae eggs.(AU)

Inmovilización química y evaluación de salud de vizcachas salvajes (Lagostomus maximus) en el chaco arido argentino / Chemical restraint and sanitary assessment of wild vizcachas (Lagostomus maximus) in the arid argentin chaco / Contenção farmacologica e avaliação sanitaria de viscachas selvagens (lagostomus maximus) no chaco árido argentino

Con el objetivo de evaluar la salud de vizcachas salvajes (Lagostomus maximus), se capturaron en el Chaco Árido argentino 10 ejemplares y se efectuaron 11 inmovilizaciones con anestesias fijas o inhalatoria. Para las anestesias, se administraron una combinación de tiletamina- zolazepam (n = 7) o una combinación de medetomidina y ketamina (n = 2) o gas isofluorano con un vaporizador portátil creando un circuito semicerrado (n = 2). Los estudios de salud comprendieron hematología, bioquímica sanguínea, análisis parasitológicos y de exposición a agentes infecciosos mediante cultivo y serología. El uso de tiletamina-zolazepam resultó en una pobre calidad anestésica y recuperaciones prolongadas. Por El contrario, el uso de medetomidina-ketamina o de isofluorano, proporcionaron inmovilizaciones más seguras. Se concluye que el protocolo testado de tiletamina-zolazepam no sería recomendable para esta especie, sobre todo en vida libre. La búsqueda de anticuerpos para los agentes infecciosos evaluados resultó negativa, por otra parte, se aisló Listeria innocua serovar 6a em hisopados rectales, se constató infestación con pulgas (Pulex irritans), ooquistes de Eimeria lagostomi, huevos de la Familia Trichostrongylidae y huevos similares a Heteroxynema viscaciae

RESUMO: Com objetivo de avaliar a condição sanitária de viscachas selvagens (Lagostomus maximus), foram capturados, no Chaco Árido argentino, 10 animais e realizadas 11 imobilizações, com anestesia injetável ou inalatória. Empregaram-se três diferentes protocolos: tiletamina-zolazepam (n = 7), metetomidina-cetamina (n = 2) ou isoflurano com vaporizador portátil em sistema semi-fechado (n = 2). A avaliação sanitária consistiu de hematologia, bioquímica sérica, exames parasitológicos e avaliação da exposição a agentes infecciosos por cultura e sorologia. O uso de tiletamina-zolazepam proporcionou má qualidade anestésica e recuperações prolongadas, enquanto a combinação medetomidina-cetamina, possibilitou imobilizações mais seguras. Assim, o protocolo testado de tiletamina-zolazepam não é indicado para esta espécie, especialmente em animais de vida livre. A pesquisa de anticorpos para os agentes infecciosos avaliados foi negativa, mas isolou-se Listeria innocua serovar 6a de suabes retais, e foram encontradas pulgas (Pulex irritans), oocistos de Eimeria lagostomi, ovos de Trichostrongylidae e ovos similares a Heteroxynema viscaciae

ABSTRACT: In order to evaluate the sanitary condition of wild vizcachas (Lagostomus maximus), 10 animals were captured in the Arid Chaco of Argentina and 11 immobilizations were performed through injectable or volatile anesthesia. Three different protocols were used: tiletamine-zolazepam (n = 7), medetomidine-ketamine (n = 2), or isofluorane with a portable vaporizer in a semi-closed circuit (n = 2). The sanitary assessment consisted of hematology, blood chemistry, parasitological analysis, and the assessment of the exposure to infectious agents by culture and serology. The use of tiletamine-zolazepam caused poor anesthetic quality and prolonged recoveries, while the medetomidine-ketamin combination, however, caused safer immobilizations. Thus, the use of the tested protocol of tiletamine-zolazepam is not advisable for this species, mainly for free-ranging animals. The search of antibodies for the evaluated infectious agents was negative, but there was isolated Listeria innocua serovar 6a from rectal swabs , fleas were found (Pulex irritans), oocytes of Eimeria lagostomi, Trichostrongylidae eggs as well as similar Heteroxynema viscaciae eggs.

Seasonal variations in the heterologous binding of viscacha spermatozoa: A scanning electron microscope study

Seasonal changes in the reproductive activity of the adult male viscacha (Lagostomus maximus maximus) were investigated during the annual reproductive cycle. Assays of heterologous in vitro binding between compatible gametes were used to evaluate the ability of viscacha spermatozoa to achieve primary binding during its annual reproductive cycle. Sperm were collected by mincing cauda epididymis in HECM-3 medium and the sperm concentration and motility were evaluated. Cumulus-free and zona-free oocytes obtained from superovulated hamsters were inseminated in vitro with capacitated sperm suspensions, incubated at 37ºC, 5% CO2 for 3 h, and then processed for studies by scanning electronic microscopy. Statistical analysis was used to compare the quantitative differences. The number of spermatozoa significantly decreases during the regression period, while sperm motility was progressive speed in both periods. During the active period elevated sperm binding to cumulus-free and zona-free oocytes was observed, while the binding during the regression period decreased drastically. In both periods, oocyte microvilli covered sperm heads and tails. These results suggest that the ability of viscacha spermatozoa to participate in gamete recognition is profoundly affected. This would likely be related to different functional stages of the spermatozoa and their epididymal microenvironment during the annual reproductive cycle of viscacha.