P.D. Lewis

Effect of thyroid deficiency on cell acquisition in the postnatal rat brain: A quantitative histological study

The mechanisms underlying transient reduction of cell number in the developing cerebellum of thyroid-deficient rats have been studied by quantitative histological methods. Thyroid deficiency has no significant effect on the generation cycle of dividing cells in either the subependymal layer of the lateral ventricular walls or the external granulay layer of the cerebellum: the length of the cell cycle and the duration of the different phases of the cycle, including the DNA synthesis time appears to be normal. However, the external granular layer of the cerebellum contains fewer cells than in control at 12 days. Pyknotic nuclei are prominent in the granule cell layer of the hypothyroid cerebellum at this age. These amount to an estimated loss of about 1% of the total cerebellar cell population in 24 h. It is suggested that death of granule cells is for the most part a consequence of reduced Purkinje cell dendritic arborization, with failure of parallel fibre/Purkinje cell synaptogenesis. In the second postnatal week, granule cell death and reduced numbers of cells in the germinal zone can account to a great extent for the observed shortfall in cerebellar DNA content. The eventual attainment of normal cell numbers in the cerebellum of hypothyroid rats is related to a persistent external granular layer in the forth and fifth postnatal weeks.

Effect of thyroid deficiency on postnatal cell formation in the rat brain: A biochemical investigation

In thyroid deficiency, interference with postnatal cell formation seems to be confined to those regions of the brain where postnatal neurogenesis is significant. In comparison with controls the increase in cell number in the cerebellum is retarded in the second week of life, but a normal number is reached by 35 days. In contrast the DNA content of the olfactory bulbs is apparently irreversibly depressed. Mitotic activity, in terms of incorporation of [2-14C]thymidine into DNA, is mainly affected in the cerebellum: in thyroid deficient rats, it is depressed below control levels at day 12, but it is about 4 times higher than in controls at day 21 when, under normal conditions, cell proliferation has virtually ceased. The time course (15-240 min) of [14C]thymidine metabolism at day 14 shows regional differences in control rats. The rate of conversion of [14C]thymidine to [14C]thymidine nucleotides, and of these in turn to [14C]DNA is slower in the forebrain - where cell proliferation occurs on a smaller scale - than in the cerebellum. Consequently, in the forebrain nearly linear DNA synthesis rate is maintained for a longer time than in the cerebellum (1 h vs. 0.5 h), and since less 14C is conserved in DNA a significant efflux of unconverted [14C]thymidine is evident during the experimental period. The effect of thyroid deficiency on [14C]thymidine metabolism in the brain is only slight, and is due to an abnormally large supply of [14C]thymidine consequent to depressed systemic utilization of this precursor.

Effects of thyroxine on postnatal cell acquisition in the rat brain

The effects of treatment with L-thyroxine (3 micrograms by subcutaneous injection daily from birth) on cell acquisition in the rat brain were studied during the first 3 postnatal weeks. In the forebrain, thyroxine has no effect on cell proliferation in the first 6 days, but it causes decreased cell acquisition from 12 to 21 days so that cell number becomes significantly reduced. Estimates of cell proliferation kinetics and of cell death in the lateral ventricular subependymal layer show no apparent abnormality. In the cerebellum, treatment from birth leads to increased cell proliferation during the first week: in comparison with controls, the rate of [3H]thymidine incorporation into DNA, thymidine kinase activity, and the number of cells both in the major germinal site (external granular layer: EGL) and in the whole cerebellum are elevated. This initial effect of thyroxine appears by day 3 and is short-lived, being no longer evident after day 6. The build-up of cell numbers in the EGL at day 6 seems to be related to a preceding, transient retardation of cell migration from this layer rather than to an acceleration of cell replication, since cell cycle parameters are normal. From day 12 the rate of [3H]thymidine incorporation into DNA is severely reduced in treated rats. Advancement of cellular differentiation rather than increased cell death in the EGL appears to be involved in this phenomenon.

PERCUTANEOUS NEEDLE BIOPSY IN THE DIAGNOSIS OF MUSCLE DISEASES

Abstract Percutaneous needle biopsy of skeletal muscle can be used in the diagnosis of muscle disease. Because the technique is easy and readily repeated it offers a number of advantages over open biopsy, including the study of the course of a disease, the response to treatment, the early diagnosis of systemic disease, and the investigation of carriers. No complications followed needle biopsies in thirty-two patients with symptoms and signs of muscle disease, and the material obtained was adequate, both in quantity and state of preservation, for examination by light and electron microscopy.

Effect of chlorpromazine on cell proliferation in the developing rat brain. A combined biochemical and morphological study

Chlorpromazine, a widely used drug in current clinical practice, produced a severe reduction of the rate of [3H]thymidine incorporation into brain DNA of 11-day-old rats. The depression of in vivo synthesis rate was detectable by 6 h after chlorpromazine administration (50 mg/kg, s.c.) and the rate was less than 40% and 60% of controls during period 14-30 h in forebrain and 6-30 h in cerebellum respectively. The depression was dose-dependent and half maximal effect was produced with about 10 mg/kg chlorpromazine. The drug caused some retardation in the rate of conversion of [3H]thymidine to [3H]thymine nucleotides in the brain, but the severe depression in DNA labelling was also evident after correcting the values on the basis of [3H]thymine nucleotides concentration. Mitotic activity was significantly reduced in the cerebellar external granular layer. Increased numbers of cell degenerations, shown by Feulgen cytophotometry to be postmitotic, were seen in both layers 12 and 32 h after chlorpromazine. Analysis of cell cycle parameters showed no significant changes. However, the labelling index in subependymal cells was reduced, indicating an increase in turnover time of about 40%. The results are consistent with an action of chlorpromazine on cell proliferation, either by direct effects on the generation and survival of cells, or via its major pharmacological actions on neurotransmitter balance. These effects are potentially of functional and clinical significance.