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Biomedical subjects

M H Hardy

Publications and source records attributed to M H Hardy.

At least 37 records · Page 2Linked to original sources
PubMed1988

An inherited connective tissue disease in the horse.

The hyperextensible, fragile skin of two related horses was compared with the skin of eight normal horses. Skin sections were examined by light microscopy and transmission electron microscopy. The deep dermal layer of the dorsal abdomen was much thinner in the affected horses, and contained bundles of collagen fibers which were more loosely packed. Within individual fibers, the fibrils were frequently curved and nonparallel rather than straight and parallel. Both of the affected animals had a greater range of fibril diameters than a normal horse. They had some unusually thick fibrils with very irregular outlines in cross-sections, not observed in the normal animal. Other skin samples were subjected to acetic acid extraction, pepsin digestion, amino acid analysis and polyacrylamide gel electrophoresis. In the skin of the two affected horses, the proportion of total extracted collagen which was acid-soluble was twice as high as in two normal horses. Collagen types I and III were present in similar proportions in normal and affected horses, and the collagen chains were of normal molecular weights. The disorder resembles the group described by Minor (Minor RR: Am J Pathol 98: 226, 1980) as 'dominant collagen packing defect I' which has been reported in dogs, mink, and cats, and which shares features with Ehlers Danlos Syndrome I, II, and III in man. The pedigree data available for these horses suggest an autosomal recessive mutation, but are also consistent with autosomal dominant inheritance.

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PubMed1986

The development of the Syrian hamster cheek pouch.

The objective of this study was to provide a detailed account of the morphogenesis and early cytodifferentiation of the hamster cheek pouch. Although the newborn "cheek pouch" is used for in vitro studies of the effects of retinoids and carcinogens, its rudimentary structure has not been adequately described. Complete paraffin serial sections of the heads of 14- and 15-day fetuses were cut in three planes to determine the location and shape of the earliest pouch rudiments. Complete paraffin serial sections were prepared from pouch rudiments dissected from hamsters at birth and at daily intervals from 3 to 12 days postnatal. Semithin Epon sections were examined by light microscopy and ultrathin sections by transmission electron microscopy. The pouch can appear in the fetus as two solid epithelial ingrowths from the lining of the oral cavity. They are the margins of an ingrowing sheet of oral epithelium which becomes leaflike at about the time of birth, as it grows caudad into the tissue of the cheek. The central cells of the ingrowth accumulate large quantities of glycogen before differentiating as a stratum spinosum 5 days after birth. Within the stratum spinosum, groups of cells containing keratohyalin granules initiate the stratum granulosum. Keratinized cells appear within the stratum granulosum areas. Spaces appear between keratinized cells, and the spaces coalesce to form the pouch cavity between 7 and 12 days postnatal. Soon afterward, this cavity opens to the oral cavity to make a pouch, and the ultrastructure of the cheek pouch epithelium closely resembles that of the adult.

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PubMed1984

The differentiation of the dermis in the laboratory mouse.

The purpose of this study was to provide a comprehensive and sequential account of the differentiation of the dermis in one body region in a mammalian species. A histological, histochemical, and ultrastructural study was made of each cellular and matrix component of the dermis of the upper lip of the mouse during prenatal development. On the basis of these observations, the development of the dermis was divided into four phases: I) undifferentiated mesenchyme (12, 13 days), II) cell differentiation (14, 15 days), III) dynamic transition (16 days), and IV) matrix differentiation (beginning at 17 days). The first phase was marked by a decrease in the cell density but no change in the ultrastructure of the undifferentiated mesenchyme cells. The second phase began with the cytodifferentiation of the mesenchyme cells and was characterized by the appearance of new cell types in the dermis (immature fibroblasts, mast cells, myoblasts, and cells of indeterminate type). During phase III the dermis was undergoing rapid change. Fibroblasts became fully differentiated, mast cell density reached a sharp peak, there was a marked increase in the number of collagen fibrils in the dermal matrix and the first collagen fibers were observed, and changes occurred in the pattern of proteoglycan synthesis. Aggregations of vesicles appeared to be extruded from cytoplasmic blebs on the fibroblasts in large quantities at this time. Further differentiation of the dermal intercellular matrix occurred during the fourth phase, which continued after birth, as more collagen was laid down to form the connective tissue stroma.

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PubMed1983

Basal lamina changes during tissue interactions in hair follicles--an in vitro study of normal dermal papillae and vitamin A-induced glandular morphogenesis.

Skin pieces from 14-day fetal mice were cultivated for 1-10 days prior to fixation and sectioning. Subsequently, sections were studied by light and transmission electron microscopy. In a standard medium the lateral hair follicle walls showed progressive maturation of the basal lamina, while the hair matrix, at the time of a known tissue interaction, showed the formation of gaps in the basal lamina, with heterotypic cell contacts through the gaps. In a vitamin-A enriched medium similar changes occurred, not only at the hair matrix, but also at lateral follicle walls, at the sites of, and prior to, budding and glandular morphogenesis. This study shows that the induction of hair matrix by dermal papilla may perhaps be added to the list of normal tissue interactions in which heterotypic cell contacts occur. It also suggests that vitamin-A induced glandular morphogenesis might come about through a mechanism resembling a normal tissue interaction.

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PubMed1983

Morphological changes at the basement membrane during some tissue interactions in the integument.

Changes at the basement membrane have been reported at the time of tissue interactions during the development of a number of integumentary organs. During the development of vibrissa follicles of the mouse in vitro, gaps appeared in the lamina densa surrounding the dermal papilla at the time of the second, specific dermal message for follicle differentiation. Direct contacts between epithelial and mesenchymal cells were made through these gaps. This area has now been studied in vivo over the same period of follicle development. In addition, the epithelial part of the follicle has been separated from the dermal papilla by (i) enzyme treatment which destroys the lamina densa and (ii) chelating agents which leave the lamina densa attached to the dermal papilla. The resulting surfaces of epithelium and dermal papilla were examined by scanning and transmission electron microscopy. Small gaps first appeared in the lamina densa at the same substage of follicle development as in vitro and were present at the next substage. Processes from the dermal papilla mesenchyme cells traversed these gaps and made close contacts with the epithelial cells just prior to their differentiation into inner root sheath and hair. Instructive interaction through cell surfaces is suggested.

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PubMed1980

A spatial relationship between innervation and the early differentiation of vibrissa follicles in the embryonic mouse.

The present study has demonstrated that the mystacial vibrissae of the mouse began to develop at about 12 days of gestation on two plates of thickened ectoderm called the 'whisker pads' which were located on either side of the snout above the margin of the upper lip. Each whisker pad was traversed by five rostrocaudal skin ridges. The individual vibrissae developed along the ridges in a caudorostral sequence. Four new sub-stages of vibrissa follicle development which occurred prior to Stage 1 of Davidson & Hardy (1952) were described. The first of these, Stage A, was the formation of a small nerve plexus under the skin ridge. Stage B was then characterized by the formation of a dermal condensation above the nerve plexus. The epithelium over the dermal condensation began to thicken at Stage C and grow down into the dermal condensation at Stage D. The early morphogenesis of the mystacial vibrissa follicles of the mouse was compared to that of teeth and mammary glands. The possibility of nerve involvement in determining the pattern of follicle array on the snout was discussed. The sequence of the morphological changes in the dermal and epidermal components of the early follicles was related to the present knowledge of epithelial-mesenchymal interactions which occur during this phase of follicle development.

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PubMed1980

Formation of feathers on chick foot scales: a stage-dependent morphogenetic response to retinoic acid.

In the chick embryo, large scales (scuta) form on days 9-11 on the anterior face of the shank and the upper surface of the toes; smaller scales (scutella) appear on day 11 on the posterior face of the shank; most of the smallest scales (reticula) form on days 12 and 13 on the plantar surface of the foot. A single injection of 125 micrograms of retinoic acid (RA) into the amniotic cavity caused the formation of feathers on the foot scales in locations and percentages differing according to the age of embryos at the time of treatment. Injection at 10 days caused feathers to form on the feet in 57% of the embryos, almost all of which bore feathers on their scuta and scutella; a few reticula were occasionally affected. When treated at 11 days, 48% of embryos had feathered feet, the tarsometatarsal scutella and digital reticula being most frequently affected. Treatment at 12 days resulted in feathers on the feet of 15% of the embryos, all of which bore feathers on the reticula only, while the scuta and scutella were not affected. It was concluded that the foot skin regions which were affected by RA treatment were those in which scale morphogenesis was starting or about to start at the time of injection. These regions contained pre-placodal or placodal stages of scuta and scutella, and the pre-elevation or elevation stages of reticula.

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PubMed1979

Localization of vtiamin A by autofluorescence during induced metaplastic changes in cultures of skin.

A technique was devised for following the uptake and location of vitamin A in organ cultures. Explants of 12- and 13-day embryonic mouse upper lip skin were grown for 3, 6 or 9 days in biological medium to which was added 0, 4.1 or 6.9 micrograms per ml of retinyl acetate. This form of vitamin A caused glandular morphogenesis of vibrissa follicles, and keratinization in epidermis and follicles was completely suppressed in 12-day explants and partially suppressed in 13-day explants. Frozen sections at 16 microns showed the white, nonfading fluorescence of keratin and the green, rapidly-fading fluorescence due to vitamin A which was captured by high-speed photography. Although more concentrated within lipid droplets in the dermis, the vitamin penetrated both the epidermis and the hair follicles. The ability to obtain permanent photographic records of the fading fluorescence makes this a useful method for analyzing vitamin A distribution as well as keratin distribution.

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PubMed1978

The stability of vitamin A-induced metaplasia of mouse vibrissa follicles in vitro.

Tests were made of the stability of the previously described glandular morphogenesis and mucous metaplasia of embryonic mouse vibrissa follicles produced in vitro by excess vitamin A. The changes in individual follicles were observed in living tissue explants and serial sections. Upper lip skin of 13.5-day embryos underwent budding from vibrissa follicles to form branching glands which secreted mucus after 10--14 days in medium containing 4.7 microgram/ml retinol. If this medium was replaced with standard medium after 7 days, glandular morphogenesis and metaplasia continued unaffected, although some hair follicle bases returned to their original morphogenetic program. Similar results were obtained with skin of 13-day embryos treated with vitamin A (6.0 MG/ML) for only 3 days. The moderate degree of glandular morphogenesis in 15-day skin with vitamin A (4.7 microgram/ml) was not altered by the simultaneous addition of a high dose of cortisol (18 microgram/ml). It is suggested that the initiation of glandular morphogenesis of follicles differs from the initiation of mucous metaplasia in embryonic chick epidermis by vitamin A, in that it resembles a new secondary embryonic induction rather than a modulation of the epithelium.

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PubMed1977

Histochemical evidence of mucosubstances in the metaplastic epidermis and hair follicles produced in vitro in the presence of excess vitamin A.

Pieces of upper lip and lower jaw from 13-day old mouse embryos grown in organ culture with excess vitamin A showed metaplastic changes in epidermis and hair follicles after 7 to 21 days. Changes were less marked in upper lip explants from 15-day old embryos. Areas of keratinizing epidermis showed a much higher incidence of Alcian blue-positive bodies when excess vitamin A was present. Histochemical tests with Alcian blue at critical electrolyte concentrations indicated moderately sulfated acidic mucosubstances in these bodies. Other patches of epidermis were transformed into stratified cuboidal epithelium producing PAS-positive, Alcian blue-negative, mucosubstances. The hair follicles, transformed by vitamin A into glands, developed three types of epithelium with apical granules, luminal borders and occasional goblet cells, all showing mucosubstances. Tests suggest that both metaplastic epidermis and glands secrete either neutral mucosubstances or non-sulfated acidic mucosubstances. Another response to vitamin A was a rapid loss of glycogen, particularly in hair follicles and the epidermal basal layer. It was concluded that excess vitamin A alters not only the morphogenesis but also the distribution and synthesis of polysaccharides in developing skin.

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PubMed1976

Ciliated and secretory epidermis produced from embryonic mammalian skin in organ culture by vitamin A.

When skin from the upper lip of 12-day embryonic mice was grown for ten days in organ culture with 5.7 mug retinol added per ml of biological medium, keratinization was suppressed and a ciliated and secretory epithelium was produced. Ultrastructural features of this epithelium are described. At this very early stage mouse epidermis is thus similar of chick epidermis in its ability to undergo radical metaplasia in response to vitamin A.

Animals↗
PubMed1976

In vitro development of secondary blastodiscs from dispersed blastoderm cells of Gallus domesticus.

A study of the in vitro growth of embryonic structures from dispersed blastoderm cells is reported. The specific type of blastoderm cells with the capability of growing on the coverslip developed into the discoidal embryonic structures resembling those of avian species. The glass surface was apparently an initiator of differentiation into at least three types of cells specifically distributed in the blastodisc. Groups of structures formed were evaluated at 6, 12, 24 and 36 hours to study the developmental pattern in vitro and to estimate the number of cells per structure. Microscopic examination of the area pellucida revealed that all three basic germ layers were established after 24 hours of incubation in vitro. The 36 hour stage was represented by bulky growth of mesodermal-like cells and changes in hypoblast layer where some of the cells degenerated and some were transformed to mesenchymal spindle-like cells.

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PubMed1975

Effects of steroid hormones on developing mouse skin in vitro.

Pieces of skin from 13-5- to 15-day-old foetal mice were grown in organ culture in a biological medium with or without the addition of hormonal steroidsmcortisol (7-5 mug/ml) caused thinning of the non-cornified epidermis and flattening of the stratum granulosum after 3 days. By 6 days the epidermis was thinner and hair follicles were regressing, and these changes continued up to 12 days. Administration of corticosterone (5 mug/ml) also produced thinning of the epidermis and regression of the follicles after 6 days. Good differentiation of epidermis and hair follicles was obtained when testosterone (100 mug/ml) was added to the medium. The non-cornified epidermal layers were similar to those of control cultures at 3 days but less than half as thick at 6 days. Hair follicles differentiated as rapidly in medium containing testosterone as in normal medium, but, unlike in the latter medium, also developed sebaceous gland anlagen at 6 days. Some explants in testosterone medium showed signs of sebaceous cell differentiation at 9 days.

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