Perinatal development of the mammillothalamic tract and innervation of the anterior thalamic nuclei.

Axonal projections originating from the mammillary bodies represent important pathways that are essential for spatial information processing. Mammillothalamic tract is one of the main efferent projection systems of the mammillary body belonging to the limbic "Papez circuit". This study was aimed to describe the schedule of the mammillothalamic tract development in the rat using carbocyanine dye tracing. It was shown for the first time that fibers of the mammillothalamic tract being the collaterals of the mammillotegmental tract axons start bifurcating from the mammillotegmental tract on E17. The axons of the mammillothalamic tract grow simultaneously and reach the ventral region of the anterior thalamus where they form first terminal arborizations on E20-E21. Ipsilateral projections from the medial mammillary nucleus to the anteromedial and anteroventral thalamic nuclei develop from E20 to P6. Bilateral projections from the lateral mammillary nucleus to the anterodorsal thalamic nuclei develop later, on P3-P6, after the formation of the thalamic decussation of the mammillary body axons. Unique spatial and temporal pattern of the perinatal development of ascending mammillary body projections to the anterior thalamic nuclei may reflect the importance of these connections within the limbic circuitry.

[Perinatal development of mammillotegmental connections in rats].

Development of direct axonal connections of the hypothalamic mammillary bodies with ventral and dorsal tegmental nuclei of Gudden was studied on fixed rat brains from day 14 of embryonic development until day 10 of postnatal development using the method of diffusion of the lipophilic fluorescent carbocyanine tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate along the neuronal membranes. The tracer was inserted into the mammillary bodies or into the tegmentum and after incubation in a fixative fluorescent nerve cells and nerve fibers were visualized in the brain tissue. The mammillotegmental tract was found to start developing earlier than other conducting systems of the mammillary bodies. On days 14-15 of embryonic development, it was visualized as a bundle of axons running from the mammillary bodies caudally to the midbrain. A group of neurons in the midbrain tegmentum and their axons going to the mammillary bodies via the mammillary peduncle were first visualized on day 19 of embryonic development. The mammillotegmental tract and mammillary peduncle developed progressively from the moment of birth. Ventral and dorsal tegmental nuclei were formed in the midbrain by day 10 of the postnatal development. Thus, the formation of reciprocal connections of the mammillary bodies with midbrain tegmental nuclei was first described during perinatal development in rats.

Immunohistochemical analyses of thyroid-specific enhancer-binding protein in the fetal and adult rat hypothalami and pituitary glands.

Thyroid-specific enhancer-binding protein (T/EBP), also known as NKX2.1 or TTF-1, regulates the expression of thyroid- and lung-specific genes. The t/ebp/Nkx2.1-null mutant mouse was stillborn but lacked the thyroid gland, pituitary gland, ventral region of the forebrain and normal lungs. These data demonstrated that T/EBP/NKX2.1 plays an important role not only in tissue-specific gene expressions in adults but also in genesis of these organs during development. Although the expression of t/ebp/Nkx2.1 in the brain has been reported, its function in the brain remains unknown. The present study was designed to determine the localization of T/EBP/NKX2.1 in the hypothalamus and pituitary gland of fetal and adult rats by immunohistochemistry as the first step toward understanding the function of T/EBP/NKX2.1 in the rat brain. In the fetal rat hypothalamus, T/EBP/NKX2.1 was localized widely in the ventral hypothalamic areas. In the adult rat brain, T/EBP/NKX2.1 was localized in the ventromedial hypothalamic nucleus, medial tuberal nucleus, arcuate nucleus and mammillary body. No T/EBP/NKX2.1 immunoreactivity was observed in the anterior or intermediate lobe of the pituitary gland in either fetal or adult rats. On the other hand, immunoreactive T/EBP/NKX2.1 was found in the posterior lobe of the pituitary gland. This paper presents results of detailed analyses of the distributions of T/EBP/NKX2.1 protein in the fetal and adult rat hypothalami and pituitary glands, and these results should provide important information for understanding the function of T/EBP/NKX2.1 in the brain.

The winged helix gene, Foxb1, controls development of mammary glands and regions of the CNS that regulate the milk-ejection reflex.

The ability to lactate is a process restricted to mammals and is necessary for the survival of nonhuman mammals. Female mice carrying a null mutation in the winged helix transcription factor Foxb1 (previously Mf3/Fkh5/TWH) have lactation defects on inbred genetic backgrounds. To determine the cellular basis of the Foxb1 lactation defect we have inserted a tau-lacZ lineage marker into the locus to follow the fate of Foxb1 expressing cells. This approach has revealed that Foxb1 is expressed in epithelial cells of developing and adult mammary glands as well as previously described regions of the central nervous system. Mammary glands from C57BL/6 Foxb1-/- mice have incomplete lobuloalveolar development. In addition, the tau-lacZ lineage label was used to determine that the mammillothalamic tract was lost in all Foxb1-/- mice. Finally, morphological defects in the inferior colliculi of the midbrain in Foxb1-/- mice correlate with the inability to lactate, suggesting that the midbrain defect, but not the loss of the mammillothalamic tract, may be responsible for the lactation defect.

Development of the mammillothalamic tract in normal and Pax-6 mutant mice.

The mammillary bodies represent important relay stations for one of the major neuronal circuits in the brain: the limbic circuit. Mammillary projections traveling through the principal mammillary tract are established early during development, forming the mammillotegmental bundle, which appears fully developed by embryonic day 15 (E15). The mammillothalamic tract develops later, around E17-E18, forming a compact system of collateral fibers originating from the principal mammillary tract and reaching the thalamus by E20. The Pax-6 gene is expressed in various regions of the developing brain, among which the border separating the ventral thalamus from the dorsal thalamus, known as the zona limitans intrathalamica, is especially significant. In this report, the development of the efferent mammillary system of fibers was studied in wild type and Pax-6 mutant mice by using carbocyanine tracers and Golgi preparations. In mutant mice, the mammillotegmental bundle developed normally; however, the mammillothalamic tract was missing. By using anti-Pax-6 antibodies in wild type mice, the existence of an immunoreactive cell cluster is described surrounding the bifurcation point of the principal mammillary tract. The results of this study suggest that there is a correlation of these cells with a particular type of Golgi impregnated neuron.

Functional sexual dimorphism of the nucleolar organizer regions in the tuberomamillary nucleus.

The tuberomamillary nucleus (TM) is a cluster of magnocellular neurons that are located close to the mamillary recess or basal surface of the mamillary body (MBs) and is the only known source of histamine (HA). The nucleolar organizer regions (NORs) of the nervous cells of the subnuclei E1, E2 and E3 of the TM are quantified in this paper to discern the existence of possible sexual differences in the overall neuronal protein synthesis between male and female rats in two phases of the oestrous cycle (oestrous-dioestrous). We have used the argentic impregnation technique of the NORs (AgNORs) in both bilateral nuclei of the TM and found functional bilateral symmetry in its AgNORs. Furthermore, we demonstrate the existence of significant differences (P < 0.001) between sexes and between two phases of the oestrous cycle studied (oestrous-dioestrous) in the parameter of number of AgNORs per neuron. Hormonal manipulation of the TM in the rat during the two periods, postnatal or adult, shows the importance of the activational effect of the hormones on the activity of cellular protein synthesis.

Postnatal development of argyrophilic nucleolar organizer regions in the mammillary body of undernourished rats.

The effects of undernutrition during pregnancy and/or lactation periods on cortical structures have already been reported. However, its effect on non-cortical areas like the hypothalamic, that participates in the regulation of the autonomic nervous system or food intake, has not been extensively investigated. We studied the postnatal development of the medial mammillary nucleus (hypothalamus) in the rat offspring that had a dietary restriction in utero and during lactation. The argyrophilic nucleolar regions (Ag-NORs), that represent the transcriptional activity of the cell, were quantified in their neurons. No statistically significant differences were found in the results between the control and undernourished groups in the ages studied (7, 14, 21 and 30 days). However, a delay in the neuronal activity of the latter group was observed. A critical period in the development of this nucleus, at about 21 days of age, was also seen in both groups. After this age, the activity levels remained steady.

Age-related shrinkage of the mamillary bodies: in vivo MRI evidence.

Mamillary body (MB) is a diencephalic structure hypothesized to be involved in memory, a cognitive function that declines with age. In this study, age-related differences in the size of MB were examined in vivo using magnetic resonance (MR) imaging. The cross-sectional area of the MB was estimated from MR images of the brain in healthy volunteers and neurologically intact patients (age 18-78). The cross-sectional area of the tectum was used as a control region of interest. We found a significant age-related reduction in the area of the MB, but not of the tectum. No sex differences were observed in the size of either structure.

Postnatal development of forebrain regions in the autoimmune NZB-mouse. A model for degeneration in neuronal systems.

NZB-mice are known to have impaired cognitive functions. The aim of the present study is the analysis of the volume growth of different brain regions in NZB/NBOM-mice, because the functional impairment increases postnatally. The regions analysed include brain structures which are important for learning and memory functions. The comparison between NZB-mice and controls (CFW- and Balb/c-mice) shows that the hippocampal volume in NZB-mice is larger than in controls. However, ectopic neurons are found in the dentate gyrus of NZB-mice, indicating a changed connectivity in this region. The septum and the amygdala show no difference in volume in NZB-mice compared to controls. The adult volume of the entorhinal cortex of the NZB-mice is the smallest of the three strains. The development of this brain region is characterized by an overshooting growth in all strains. The caudate-putamen complex and the globus pallidus of NZB-mice undergo a reduction in volume during the postnatal period. This is not found in the controls. An overshooting growth is seen in the mamillary bodies of the three strains, and in the anterior thalamic nucleus of NZB-mice. However, only the NZB-mice show a prolonged reduction of the volume of the mamillary bodies, which is not finished during the observed time period. Both regions are important relay stations in the Papez-circuit, a neuronal system associated with learning and memory functions. The prolonged postnatal reduction in volume of the mamillary bodies and the anterior thalamic nucleus of NZB-mice may be the structural correlate of the impaired cognition and memory in this strain.

[Quantitative observations of non-neuronal and neuronal cells of the mammillary bodies during postnatal development]. / Observaciones cuantitativas sobre las células no neuronales y neuronales de los cuerpos mamilares durante el desarrollo postnatal.

The mammillary bodies are already shaped in the last stages of the fetal development, and you can differentiate clearly in them the medial mammillary nucleus (MMN) and lateral mammillary nucleus (LMN). All this allow to study the variations in cellular density, the relation between neuronal and glial cells, so as the number of nucleoli present in the neuronal cells and the nucleolar area variations through the postnatal development (0N, 2N, 4N, 8N, 16N and adult). The results show, demonstrate a different behaviour in the cellular stabilization and migration of the MMN and LMN, so as a cellular differentiation after 4N in both nucleus. All this is discussed with the data obtained from the nucleolar activity, as an index of synthesis and reflection of the neuronal maturing.

Neurons containing cholecystokinin mRNA in the mammillary region: ontogeny and adult distribution in the rat.

1. The ontogeny and adult distribution of neurons containing cholecystokinin (CCK) mRNA in the premammillary and mammillary nuclei and supramammillary region of the rat brain were studied using hybridization histochemistry. 2. The earliest detection of CCK mRNA in the mammillary region was on E14, followed by a marked increase in transcript levels during the next 4 days, a time during which neurons in this region still divide. During the first 2 weeks of life, few changes in the levels of CCK transcripts were seen, and an adult-like pattern of expression was seen on the twenty-first day of life. 3. Low levels of transcripts were present in numerous neurons located in all divisions of the medial nucleus and in the posterior nucleus known to project ipsilaterally to the anteroventral and anteromedial thalamic nuclei. In contrast, none of the neurons in the lateral nucleus (projecting bilaterally to the anterodorsal thalamic nucleus) had detectable transcripts. 4. Many neurons in the supramammillary nucleus had low, moderate, or high levels of transcripts. Some nearby nuclei (such as the dorsal premammillary nucleus) had smaller numbers of neurons with low levels of CCK mRNA, whereas others (such as the ventral premammillary nucleus) had none.

A quantitative histological study of the anterodorsal thalamic nucleus and the lateral mammillary nucleus of ageing mice.

The number of neurons and neuroglia in the anterodorsal nucleus of the thalamus and in the lateral mammillary nucleus of the ASH/TO strain mouse was estimated at 6, 25, 28 and 31 months of age. There was no significant variation in neuron number in either nucleus with age but there was a statistically significant increase (approximately 20%) in the number of neuroglia in the anterodorsal nucleus between 28 and 31 months of age which raised the glia to neuron ratio from 0.59 to 0.72. There was no change in neuroglial number in the ageing lateral mammillary nucleus. Neurons in both nuclei lost Nissl substance. This was first evident at 28 months in the anterodorsal nucleus and at 31 months in the lateral mammillary nucleus. Lipofuscin accumulation was common in the perikarya of anterodorsal nuclear neurons from 25 months onwards but was very rare in neurons of the lateral mammillary nucleus even at 31 months.

Neurons in the rat subiculum with transient postmamillary collaterals during development maintain projections to the mamillary complex.

We recently found that in developing rats large numbers of fibers extending through the fornix initially grow well past the mamillary bodies and into the mesencephalic and pontine tegmentum (Stanfield et al. 1987). This postmamillary component of the fornix is essentially completely eliminated during the first few postnatal weeks, although the cells of origin of this projection within the subicular complex of the hippocampal region persist. To determine if the subicular neurons which transiently extend post-mamillary axons maintain a projection to the mamillary complex after the elimination of the postmamillary component of the fornix, we employed a delayed double-retrograde labeling paradigm. The cells of origin of the postmamillary component of the fornix in rat pups were labeled with an injection of fast blue (FB) into the mesencephalic/pontine tegmentum, and, in the same animals, but at a stage after the elimination of the postmamillary component of the fornix, the cells of origin of the definitive fornix were labeled with an injection of diamidino yellow (DY) aimed at the mamillary complex. In these cases the FB labeled neurons within the subicular complex can be DY labeled as well, indicating that subicular neurons which transiently extend post-mamillary axons maintain projections to the definitive targets of the fornix within the caudal hypothalamus.

[Quantitative growth analysis of limbic nuclei areas fresh volume in diencephalon and mesencephalon of an albino mouse ontogenic series. II. Corpus mammillare]. / Quantitative Analyse des Wachstums der Frischvolumina limbischer Kerngebiet im Diencephalon und Mesencephalon einer ontogenetischen Reihe von Albinomäusen. II. Corpus mammilare

The fresh volumes of the Nucl. medialis and of the Nucl. lateralis corporis mammillaris of 52 white mice aged between 17 and 60 days of ontogenesis have been determined. The fresh volume of the left Nucl. lateralis is significantly larger than that of the right side. The data of the Nucl. laterales have been fitted by a 3parametric logistic function. The data of the Nucl. medialis have been fitted by a sum of logistic functions. This sum has an increasing and a decreasing component and it shows a maximum of the sum of both components near 32 days of ontogenesis. The degree of maturity and the growth-rate of maturity have been discussed. Both nuclei belong to the group of brain regions in white mice with an early development. The growth of different brain regions is asynchronous. There is a need for further quantitative analyses.