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Biomedical subjects

G Gopinath

Publications and source records attributed to G Gopinath.

At least 19 recordsLinked to original sources
PubMed1998

Recovery of sleep after fetal preoptic transplantation in medial preoptic area-lesioned rats.

Changes in sleep after fetal preoptic (POA) tissue transplantation were studied in rats which had been made insomniac by a medial preoptic area (mPOA) lesion. Two days after the N-methyl D-aspartic acid (NMDA) lesion of the mPOA, fetal POA tissues (obtained from 14- to 17-day-old fetuses) were transplanted into the lesioned mPOA. Insomnia was less marked in these animals, as compared to nontransplanted lesioned rats, even on the 4th day after transplantation. The quantum of sleep nearly attained the prelesion level by the 20th day. Body weight also showed recovery after transplantation. Rectal temperature, which was increased by the lesion of the mPOA, remained unaltered even after the transplantation. These results suggest that the recovery of sleep and rectal temperature may follow different time courses. Surviving transplanted neurons were seen at the site of lesion on postmortem examination. Humoral interaction between the host and the transplant may be responsible for the early recovery of sleep, though the establishment of neural connections between the host and transplant might have contributed to the later recovery. This is the first study to show the recovery of sleep function in insomniac animals after fetal preoptic tissue transplantation. However, the specificity of the POA fetal tissue, in comparison with other neural tissues to promote sleep recovery, remains to be established.

Animals↗
PubMed1996

Cell surface molecules (NCAM and L1) in intrastriatal transplants of embryonic mesencephalon in rats.

Cell surface molecules, NCAM and L1, reported to have a role in synaptogenesis, growth and fasciculation of the neurites in the brain, were traced in the embryonic nigral transplants in the host striatum of adult rats. Substantia nigra of five, 15 and 25 postnatal days were also examined for the same molecules. Tyrosine hydroxylase label was used as a marker to localize the nigral neurons and glial fibrillary acidic protein to detect if glial scar present. In the control as well as transplants large neurons had expressed tyrosine hydroxylase. By 15th postnatal day tyrosine hydroxylase neurons appeared mature and were scattered, suggesting a well-formed neuropil. NCAM and L1 reaction was seen as a peripheral rim in most of the cells on the fifth postnatal day. The reaction was mainly in relation to the large cells and more extensive on the 15th day. Thereafter on the 25th day, activity was negligible. Large neurons demonstrated strong reactivity for NCAM and L1 during early post-transplantation days. After 30 days only smaller cells were reactive, many of which could be identified as neurons. Strong reaction for these molecules was present only until 60 days, though faint reaction could be detected even on the 90th day. These observations indicate that the growth promoting molecules, the type seen in the neonatal period, can be detected normally only until the neurons mature. Prolonged expression of these molecules by the grafted neurons indicate delay in the maturation of these cells due to absence of adequate target sites for synaptic connections. Some of the smaller cells expressing these molecules after 30 days of transplantation could be astroglia, either proliferating or reactive.

Animals↗
PubMed1996

Ultrastructure of developing substantia nigra in humans.

Electron microscopy of the maturing neurons and developing and maturing synapses in the substantia nigra of 14 human embryos/foetuses of 8-24 weeks of gestation are reported. At 8 weeks, cells were immature with very little cytoplasm and cellular organelles. Contact sites of processes appeared more electron dense than the other areas. At 12 weeks, many of the cells had acquired more cytoplasm and cellular organelles and could be identified as neurons. Asymmetric synapses with clear, round synaptic vesicles also were identifiable at this age. Such synapses, first to appear in the developing substantia nigra, are reported to be formed by recurrent collateral nigro-striatal fibres. Substance P fibres from the striatum also are contributing to this type of synapse. At 15-16 weeks, not only was the number of such synapses increased, but many appeared morphologically mature. Symmetric synapses having clear round vesicles along with a few dense core vesicles also appeared at this stage, suggesting striatal input. By 24 weeks of gestation, most of the neurons had cytological features comparable to that of the mature neurons. There was an increase in the total number of synapses and the individual variety from 15 to 24 weeks of gestation. The present study indicates that synaptogenesis starts at 8 weeks and continues beyond 24 weeks of gestation.

Cytoplasm↗
PubMed1996

Long-term nigral transplants in rat striatum: an electron microscopic study.

The substantia nigra of gestation day 14 was transplanted into the striatum of 3-4-month-old rats to investigate the transplants ultrastructurally at the end of 2 years, as a follow-up to our previous studies. Transplants were of small size in all 10 specimens taken for this study. The changes observed in the transplant and in the interface region with the host striatum were: thickening of the blood vessel walls, perivascular cuffing with lymphocytes and macrophages loaded with tissue debris, degenerating neurons and hypertrophied astroglia containing dense granules indicating ageing or reaction to degeneration and glial processes. The number of surviving neurons in the transplants was small. These were smaller in size and had very few intracytoplasmic membraneous organelles. A higher content of intracytoplasmic ageing lipofuscin pigment was present than in host neurons and age-matched nigral neurons. Synapses were few, and their number varied among transplants. Generally, the synapses were at the interface with the host tissue. The changes observed in all the 2-year-old transplants suggest premature ageing or a slow rejection process. Slow rejection is a possibility, because these rats are only stock-bred, not inbred, and hence they are not completely immunologically compatible.

Animals↗
PubMed1994

Developing substantia nigra in human: a qualitative study.

Midbrains from 43 fresh human embryos and fetuses at 8-22 weeks of gestation were processed for routine histology, Golgi staining, tyrosine hydroxylase (TH) immunolabelling and retrograde tracing with the fluorescent dye DiI. Cells were immature and densely packed between 8 and 10 weeks. By 13 weeks cells could be identified as neurons and glia. Neurons matured gradually and achieved adult characteristics by 20-22 weeks. Neurons in the paramedian regions of the tegmentum, raphae region and substantia nigra were positive for TH from 13 weeks onwards, the earliest age group used for this technique. The presence of TH-positive neurons in the paramedian part of the tegmentum until 18 weeks and radial glial fibers extending from the aqueductal lining to the ventral part until 20 weeks were suggestive of migration of neurons to the ventral mesencephalon region. DiI labelling of the neurons and fibers of ipsilateral nigra from the caudate as early as 10 weeks demonstrated early nigrostriatal connections. The mature nature of the neurons appeared only by 13 weeks by this method. The present study shows that the nigral neurons in the human migrate and mature until mid-gestation. The nigrostriatal connection at 10 weeks suggests a trophic influence of nigra on the proliferating and maturing neurons of the striatum.

Carbocyanines↗
PubMed1994

Changes in sleep-wakefulness after kainic acid lesion of the preoptic area in rats.

The role of the preoptic area (POA) neurons in the regulation of sleep-wakefulness (S-W) has been investigated in this study. The cell-specific neurotoxin, kainic acid (KA), was injected (0.8 microgram in 0.2 microliter) intracerebrally for lesioning of the POA. S-W was assessed (on the basis of EEG, EMG, and EOG recordings) for a day before bilateral lesion of the POA, and for 3 weeks after the lesion. There was an increase in wakefulness, and a decrease in all the stages of sleep after KA lesion of the POA. The reduction in deep slow wave sleep (S2) and REM sleep (PS) were more marked than light slow wave sleep (S1), and these had not shown any recovery even after 3 weeks of lesion. Two days after the lesion, the reduction in sleep was much more marked during the daytime than at night. There was an increase in locomotor activity, especially during the daytime, though it was only statistically significant on the 6th and the 10th day after the lesion. This study shows that the POA neurons are involved in the induction and maintenance of sleep. The lesion did not have a long lasting effect on the circadian distribution of sleep but the changes in locomotor activity seem to persist for a longer period.

Animals↗
PubMed1992

Developing dentate nucleus in man: a qualitative and quantitative study.

Histological and quantitative studies were carried out in dentate nucleus (DN) of 14 human fetuses of gestational ages ranging from 10 weeks to 28 weeks in Nissl stained sections. The dentate nucleus was already well delineated at 11 weeks, but the adult profile was attained only by 27-28 weeks. The magnocellular dorsomedial region and the parvocellular ventrolateral region became apparent at 17-18 weeks. The numerical density of neurons showed a steady decline with advancing gestational age. However, the absolute neuronal count until 19-20 weeks was 367.1 x 10(3) followed by a count of 249.5 x 10(3) at 22-23 weeks. The reduction in the count between the two periods was significant and coincided with cell death which was striking between 19 and 23 weeks. The glial cell population continued to increase with advancing gestational age. The occurrence of significant neuronal cell death and its probable role in the developing human DN is reported for the first time.

Cell Count↗
PubMed1991

Fetal nigral grafts in the anterior eye chamber of adult rats: a long-term morphological study.

Substantia nigral grafts of 16 and 17 gestation days showed phenotypic characteristics in the anterior eye chamber of adult rats until the third month after transplantation. Thereafter by the sixth month a number of neurons showed somal and dendritic thickening, reduced population of endoplasmic reticulum, increase in lysosomes, and clear areas devoid of organelles, indicating age changes. These changes were progressive and affected more neurons by the end of 1 year, the longest period studied. The observations suggest that the maturation of nigral neurons is independent of specific afferent input, whereas target influence is necessary for the continued maintenance of the mature neurons. All the synapses observed in the transplant were of the asymmetric variety, reminiscent of the few intrinsic synapses of the intact nigra. This suggested establishment of mutual connectivity among the transplanted neurons in the absence of a target and the type of synapse formed may have been influenced by the local environment. Large glial processes, very prominant during the 4- to 6-month period became less significant afterward but continued to be present until the end of the period studied. Though there was no morphological evidence of lymphocytic infiltration, this might suggest an immunologic reaction.

Aging↗
PubMed1991

Ageing changes in the transplants of fetal substantia nigra grafted to striatum of adult rat.

Fetal nigral neurons from 16 and 17 gestational days were transplanted into the intact striatum of adult rat. On different post-transplantation days (30-360 days), the structural and immunohistochemical details of the transplants were studied. The grafted neurons matured and showed phenotypical characteristics comparable to that of normal nigral neurons in adult rats until 180 days. Tyrosine hydroxylase-positive neurons were seen not only in the transplant but also in the adjacent host striatum. Tyrosine hydroxylase-positive fibres were also seen extending for a short distance into the host striatum. A large number of synapses in the transplants were of asymmetric type, containing clear round vesicles. These synapses resembled the few intrinsic type present in the normal substantia nigra. On the other hand, the predominant type 2 synapses with pleomorphic vesicles in the normal nigra were infrequently encountered in the transplants. On the 300th day, the cytoplasm of a few of the neurons showed ageing changes in the form of clear spaces, paucity of organelles especially rough endoplasmic reticulum, membrane-bound vacuoles and increase in the lipofuscin population. In addition, localized thickening of the soma and the dendrites were seen in relation to randomly distributed neurons. By 360 days, more than one quarter (26%) of the total neurons showed these changes indicating ageing. The number per unit volume of normal neurons decreased significantly when compared to the transplants on 60 and 90 days. In the substantia nigra of age-matched control, except for an increase in the lysosomal population, other ageing changes were not detectable. The neurons of intact substantia nigra of the host rat, chronologically 4-8 months older than the transplanted neurons, also appeared normal but for lipofuscin granules. The present study provides morphological evidence for rapid ageing of neurons in the long term nigral transplants. These observations raise fresh doubts regarding permanent survival of grafted neurons in the host brain. Studies so far conducted are after prior nigral lesions. Trophic factors following lesions of the host tissue may have influenced the long term survival of the transplanted neurons. On the other hand, such changes may have been missed since no detailed morphological investigations of the long term transplants have been done so far.

Aging↗
PubMed1991

Adrenal medullary autografts in anterior eye chamber, lateral ventricle and striatum of adult rats: a long term study.

The neurobiological basis for the short-term recovery in Parkinson's patients and experimental animals grafted with adrenal medulla is not yet clear. Structural details of the grafted chromaffin cells are also not available. In the present study, autografts of adrenal medulla in the anterior eye chamber, lateral ventricle and striatum of adult rats were studied for 360 days. Though a large number of cells degenerated, a few healthy chromaffin cells survived up to 360 days in the anterior eye chamber. In the ventricular and striatal regions, cells degenerated more rapidly, and a few surviving cells were seen only up to 120 and 150 days, respectively. Degeneration of the cells was evident from the alteration of the cytoplasmic granules, appearance of vacuoles and lysosomes, rapid decline in the number of TH and DBH positive cells and diffusion of enzymes in the intercellular region. Only lymphocytes and connective tissue cells were seen in the ventricle after 120 days, while outlines of a few chromaffin cells and background fluorescence were still evident in the striatum up to 150 days. In some of the intrastriatal transplants, morphologically identifiable Schwann cells were present and, in one transplant, there was evidence of myelination of axons by Schwann cells. These axons were obviously derived from the adjacent host tissue. From the findings it has been concluded that autografts of adrenal medulla survive for only a limited period of time on transplantation to the central nervous system and anterior eye chamber. Survival seemed to be better in the anterior eye chamber than at the sites preferentially chosen for treating Parkinson's patients or experimental animals.

Adrenal Medulla↗
PubMed1991

Fetal nigral transplants in the lateral ventricle of adult rats. A long-term morphological study.

The substantia nigra from rats of 16 and 17 gestational days grafted into the lateral ventricle of adult rats grew into isolated or partially integrated transplants in 80% of the rats. The morphological details of the transplants were studied at intervals of 30-360 days. The neurons grew, differentiated and were comparable to age-matched controls until 150 days. Thereafter, the neurons showed progressive aging changes like dendritic thickening, clear spaces, membrane-bound vacuoles and increase in lysosomes in the cytoplasm. The numerical density per unit volume (Nv) of normal neurons significantly decreased with simultaneous increase in Nv of glial cells by 360 days. These changes were slower to develop in the partially integrated transplants: it is concluded that target tissue interaction is necessary for prolonged survival of the grafted tissue.

Aging↗
PubMed1987

Effect of internal irradiation on the maturing Purkinje cells in the rat. A Golgi study.

Continuous irradiation in utero is reported to produce mental retardation and gross abnormalities of the brain in the human. A few experimental studies conducted so far also report gross brain defects in animals exposed to continuous irradiation in utero. Despite the increasing use of nuclear energy for power and radioisotopes in medicine, there is hardly any literature available on the effect of continuous irradiation on the structural details of the developing brain. After intraperitoneal injections of different doses of 131I (8, 18 and 32 microCi) and 32P (10 microCi) in new-born rats on the 6th postnatal day, cerebella stained by Golgi techniques were cut sagittally and the sections were examined on the 10th, 15th and 21st postnatal days. In the animals injected with 18 and 32 microCi of 131I and 10 microCi of 32P a large number of Purkinje cells showed morphological alterations not seen in the control groups or in the groups injected with 8 microCi of 131I. The changes observed included persistence of the perisomatic processes beyond the 10th postnatal day, multiple primary dendrites, angulation of the primary dendrites, long segments of primary dendrites without branches and significantly reduced dendritic volume. The number of affected cells was less on the 21st postnatal day. The effective radiation dose estimated in these groups ranged from 15 to 26 rad. Since the rats irradiated with 6 rad had not shown such changes it is believed that there is a threshold dose of radiation beyond which only changes are perceptible at neuronal level.

Age Factors↗
PubMed1984

Effects on pregnancy in mice of passive immunization against ovine LH and human chorionic gonadotrophin.

Mice given daily i.p. injections of immunoglobulins against ovine LH on Days 3-7 of pregnancy were devoid of implantation sites on Day 8 whereas mice treated with antibodies to hCG had embryos of normal number and appearance on Day 8. These antibody treatments reduced the mean +/- s.d. serum progesterone concentrations from 65.4 +/- 15.3 ng/ml (control globulins) to 8.6 +/- 4.9 ng/ml (anti-LH) and 9.2 +/- 3.1 ng/ml (anti-hCG) on Day 8 and had no differential effect on serum oestrogen levels on Day 4. However, the mice treated with anti-hCG did not litter; resorption of the embryos took place between Days 10 and 14 of pregnancy. Indirect immunofluorescence and quantitative immunoenzymic assays showed the presence of anti-ovine LH and anti-hCG reacting antigens in the mouse feto-placental unit. On Day 6, the values of reacting antigens (mean +/- s.d. absorbance units/10 micron section of embryo) were 0.050 +/- 0.002 with control globulins, 0.059 +/- 0.002 with anti-hCG-Ig and 0.196 +/- 0.018 with anti-LH-Ig; the corresponding values on Day 12 were 0.075 +/- 0.009, 0.402 +/- 0.02 and 0.416 +/- 0.015. The quantitative disposition of the reacting antigens to the two types of anti-gonadotrophins seems to bear a temporal relationship to their respective antifertility action. The pregnancy terminating action of immunoglobulins to ovine LH (Days 6, 7 & 8) and hCG (Days 8, 9 & 10) was counteracted by administration of 2 mg medroxyprogesterone acetate on Days 6, 9 and 12, indicating the importance of progesterone in the maintenance of pregnancy in the mouse.

Animals↗