A new technique for dissociation of hair follicles into single cells.

A method is described for the complete dissociation of mouse hair follicles into a suspension of single cells suitable for cell culture. Dermal tissue containing hair follicles is digested sequentially with trypsin and chondroitinase ABC under mild conditions.     

Neogenesis of functional hair follicles in adult mouse skin selectively induced by tumour-promoting phorbol esters

Summary Neogenesis of functional hair follicles in the tail skin of adult mice can quantitatively be demonstrated after long-term treatment with tumour-promoting phorbol esters. The ability to induce the formation of new hair follicles correlates with the hyperplasiogenic and tumour-promoting capacity of the phorbol esters. Hyperplasiogenic but nonpromoting phorbol esters do not lead to the formation of new hair follicles.Acknowledgment. I am grateful to Dr K. Goerttler and Dr F. Marks for helpful advice and discussions.     

Neogenesis of functional hair follicles in adult mouse skin selectively induced by tumour-promoting phorbol esters

Neogenesis of functional hair follicles in the tail skin of adult mice can quantitatively be demonstrated after long-term treatment with tumour-promoting phorbol esters. The ability to induce the formation of new hair follicles correlates with the hyperplasiogenic and tumour-promoting capacity of the phorbol esters. Hyperplasiogenic but nonpromoting phorbol esters do not lead to the formation of new hair follicles.     

Modeling melanoblast development

Melanoblasts are a particular type of cell that displays extensive cellular proliferation during development to contribute to the skin. There are only a few melanoblast founders, initially located just dorsal to the neural tube, and they sequentially colonize the dermis, epidermis, and hair follicles. In each compartment, melanoblasts are exposed to a wide variety of developmental cues that regulate their expansion. The colonization of the dermis and epidermis by melanoblasts involves substantial proliferation to generate thousands of cells or more from a few founders within a week of development. This review addresses the cellular and molecular events occurring during melanoblast development. We focus on intrinsic and extrinsic factors that control melanoblast proliferation. We also present a robust mathematical model for estimating the doubling-time of dermal and epidermal melanoblasts for all coat color phenotypes from black to white.     

Alloxan-induced hyperglycemia increases progestin and androgen accumulation by isolated rabbit follicles in vitro

Summary Follicles isolated from Alloxan-treated rabbits and incubated in vitro, accumulated more progesterone and testosterone than those from saline-treated rabbits. LH augmented the accumulation of these 2 steroids. By contrast, the estradiol response to LH stimulation by follicles from Alloxan-induced hyperglycemic rabbits was diminished when compared to follicles from saline-treated rabbits. Ovaries from hyperglycemic rabbits also appeared to have more cystic follicles.This work was supported by the Medical Research Council of Canada. The technical assistance of Misses P. Dimond and J. Osoko is gratefully acknowledged. Dr B.R. Bhavnani provided the stimulus for this study.     

Alloxan-induced hyperglycemia increases progestin and androgen accumulation by isolated rabbit follicles in vitro

Follicles isolated from Alloxan-treated rabbits and incubated in vitro, accumulated more progesterone and testosterone than those from saline-treated rabbits. LH augmented the accumulation of these 2 steroids. By contrast, the estradiol response to LH stimulation by follicles from Alloxan-induced hyperglycemic rabbits was diminished when compared to follicles from saline-treated rabbits. Ovaries from hyperglycemic rabbits also appeared to have more cystic follicles.     

Autoradiographic analysis of FSH binding during follicular atresia

Binding of FSH to ovarian cells was studied in PMSG primed immature Swiss mice, 48 h after PMSG treatment, FSH-binding was higher in the periphery than in the cumulus cells of the antral follicles. Binding of FSH to granulosa cells of normal follicles was observed to be specific, 48 h after PMSG injection. No localization in the atretic follicles could be seen by autoradiography 72 h after priming.     

Immunization of female rabbits against testosterone stimulates testosterone accumulation by isolated ovarian follicles

Ovarian follicles isolated from female rabbits after active immunization against testosterone-3-oxime bovine serum albumin produced more testosterone than similar follicles from controls.     

Excess bioavailability of zinc may cause obesity in humans

The body weight or body weight/height ratio exhibits a positive linear correlation to hair zinc (Zn) concentration which is more strongly positive in men than in women. The obese of both sexes possess higher Zn concentrations in their hair than those with normal body weight/height. The degree of obesity increases with the increase in the hair Zn concentration.     

Studies on the thyroid in transgenic mice expressing the genes for human and bovine growth hormone

The thyroid glands of transgenic mice (TM) expressing the genes for human (h) and bovine (b) growth hormone (GH) were studied. The percentages of larger follicles in hGH TM of either sex were significantly greater than in the corresponding normal littermates, and follicles ranging up to 350 microns in diameter were present in male hGH TM. In contrast, thyroid follicles were only slightly enlarged in male bGH TM, and were unchanged in female bGH TM. The serum concentrations of T4 were significantly decreased in male bGH TM and not altered in the other groups. Serum concentrations of T3 were slightly, but significantly increased in female hGH TM and female bGH TM, but were unaffected in male TM of either type. Since the principal difference between these foreign GHs in rodents is the additional lactogenic activity of human GH, these results may indicate that the effects of prolactin can influence the development of the thyroid.     

Immunization of female rabbits against testosterone stimulates testosterone accumulation by isolated ovarian follicles

Summary Ovarian follicles isolated from female rabbits after active immunization against testosterone-3-oxime bovine serum albumin produced more testosterone than similar follicles from controls.This work was supported by the Medical Research Council of Canada MT 4192.     

The inner ear structures of the echidna—An SEM study

Summary Surface structures of the echidna cochlea were investigated using a scanning electron microscope technique. It was found that unlike typical mammalian cochleas, the echidna cochlea possesses four rows of inner hair cells and 6–9 rows of outer hair cells, and that the arrangements of the stereocilia of the outer hair cells were not uniform throughout the length of the basilar membrane.We are grateful for the valuable advice given by Mr J. Nailon.     

Genetics of early mammalian folliculogenesis

Early ovarian folliculogenesis begins with the breakdown of germ cell clusters and formation of primordial follicles. Primordial follicles are the smallest ovarian follicle units continuously recruited to grow into primary and more advanced ovarian follicles. Genes expressed in the germ cells such as Figla, Nobox, Kit and Ntrk2, as well as genes expressed in the surrounding somatic cells such as Foxl2, Kitl and Ngf, play critical functions during early folliculogenesis. Transgenic mice continue to provide important insights into the genetic pathways that regulate early mammalian folliculogenesis. Genes critical in early folliculogenesis are important determinants of reproductive life span and represent candidate genes for human ovarian failure. Received 25 August 2005; received after revision 18 October 2005; accepted 21 November 2005     

Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pig ovaries

The porcine antral follicles, 3–6 mm in diameter, were dissected from the ovaries of mature pigs, and then granulosa and cumulus cells were isolated from each follicle. In atretic follicles, high activity of neutral Ca²⁺/Mg²⁺-dependent endonuclease and DNA ladder formation, estimated by electrophoresis, were noted in granulosa cells but not in cumulus cells. Extremely low activity of the endonuclease and no DNA ladder formation were observed in both types of cells obtained from healthy follicles. Moreover, apoptotic cells were observed histochemically among granulosa cells only. A good correlation (r=0.987) between the endonuclease activity of granulosa cells and the progesterone/estradiol ratio of follicular fluid in each follicle was found. These results suggest that apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pigs.     

Erythropoietin but not VEGF has a protective effect on auditory hair cells in the inner ear

It has recently been shown that the oxygen-regulated factors erythropoietin (Epo) and vascular endothelial growth factor (VEGF) confer protection on different cells, including neuronal-derived ones. The receptors for Epo and VEGF are widely expressed in different organs. Since mammalian auditory hair cells can irreversibly be damaged by different agents, we aimed to identify otoprotective compounds. We focused on the role of Epo and VEGF in the inner ear and review the recent studies. Epo and its receptor are expressed in the inner ear. In vitro experiments on auditory hair cells showed a protective effect of Epo in ischemia- and gentamicin-induced hair cell damage. In contrast, an in vivo study using an animal model of noise-induced hearing loss showed a negative effect of Epo. Also VEGF and its receptors are expressed in the inner ear. Changes in the expression of VEGF or its receptors have been found in the cochlea after noise exposure, transcranial vibration and diabetic or aged animals. Until now, there are no studies about a direct effect of VEGF on auditory hair cells in vitro or in vivo. We could exclude a protective effect of VEGF on gentamicin-induced auditory hair cell damage in vitro. Thus, we conclude that Epo but not VEGF has a protective effect on auditory hair cell damage at least in vitro. (Part of a multi-author review.)     

Sensitivity of follicular melanoblasts in newborn mouse skin to tritiated thymidine: Evidence for a long term retention of label

Summary Injection of tritiated thymidine into newborn mice results in a progressive greying of hair that does not begin until after the first hair coat is grown. After a year the depigmentation is appreciable (about 60% of the hair are white). The effect cannot be simulated by external irradiation of newborn mice or by the administration of radioactive uridine or methionine. The effect can best be explained by a long-term retention of radioactivity in the DNA of melanocyte stem cells (melanoblasts) in spite of several rounds of cell division. This could be achieved by labelling the strands of DNA destined to act as templates throughout life by being selectively retained in the stem line as described in Cairns' hypothesis.Acknowledgments. This work was supported by grants from the Cancer Research Campaign. I am indebted to Joan Bullock for her careful, patient help with these experiments. CSP is a fellow of the Cancer Research Campaign.     

Über die morphologischen Veränderungen am menschlichen Ovar unter Einwirkung eines hormonalen Antikonzeptivums

Summary Under the effect of Anovlar®, which had been administered during two cycles, one Corpus luteum per cycle developed; characteristic of each was the thecahypoplasia. The typical epitheloid Theca interna cells were lacking in the Theca interna of the existing vesicular follicles. Theca interna growth in the atretic vesicular follicles could not be demonstrated.     

Electrical resistance and spike activity in tarsal chemosensilla ofPhormia regina (Meig.)

Summary The relationship between hair electrical resistance and responsiveness to stimulation has been investigated in the tarsal chemosensilla ofPhormia regina. Results showed that the lower the hair electrical resistance, the higher is the spike-firing frequency.This study was partly supported by a grant from the Consiglio Nazionale delle Ricerche (CNR, Rome, Italy).     

Hair in Dominican amber: Evidence for tertiary land mammals in the Antilles

Summary A piece of amber from the Dominican Republic contained approximately 50 strands of mammalian hair. Based on its characteristics and the identification of two species of fossilized ectoparasites that were also present, it is probable that the hair belonged to a rodent. This find represents the earliest fossil remains of land mammals in the Antilles and lends support to the vicariance model of West Indian biogeography.     

Abnormal organization of ovarian follicles in the mutantl(1)su(f) mad-ts ofDrosophila melanogaster

Summary The morphology of ovarian follicles in the temperature-sensitive female-sterile mutantl(1)su(f) ^mad-ts has been studied by means of light and electron microscopy. As the follicle cells gradually degenerate at the restrictive temperature (29°C) the follicles become increasingly disorganized with respect to shape and polarity.Acknowledgments. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 46). We wish to thank Mrs R. Koppa for excellent technical assistance and Mrs G. Mahlke for photographic work. We are also indebted to Dr D. Zissler, Dr E. Gateff and Prof. K. Sander for many helpful suggestions.