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J D Penschow

Publications and source records attributed to J D Penschow.

29 records · Page 2Linked to original sources
PubMed1987

Alpha-inhibin gene expression occurs in the ovine adrenal cortex, and is regulated by adrenocorticotropin.

Inhibin is a glycoprotein hormone composed of two nonidentical subunits. It is produced by the ovary and testis and plays a vital role in gonadal function by inhibiting the secretion of FSH. More recently, additional activities associated with inhibin peptides have been identified. Inhibin heterodimers (alpha-beta) are reported to act directly on ovarian granulosa cells and inhibit estrogen production induced by FSH. Furthermore, homodimers of beta-inhibin subunits stimulate the secretion of FSH, an activity that is directly opposite to that of inhibin. Each of these inhibin-related activities are concerned with the hypothalamic-pituitary-gonadal axis. We have investigated further the complexity of inhibin activity by determining whether inhibin genes are expressed in nongonadal tissue. RNA hybridization experiments demonstrate that the alpha-inhibin gene is expressed in the sheep adrenal cortex and hybridization histochemistry shows that this gene is expressed in each of the functional zones within the cortex. Dot blot analysis showed that the level of alpha mRNA within the adrenal is influenced by ACTH, one of the major regulators of adrenal cortex function. These observations imply that there are inhibin-related peptides not directly associated with the gonads. beta-inhibin gene expression was not clearly detected in the adrenal and we conclude that if expression occurs then it does so at extremely low levels.

Adrenal Cortex↗
PubMed1987

Histochemical localization of IGF-I and -II mRNA in the developing rat embryo.

We describe the histological localization of embryonic and fetal tissues whose cells express the genes coding for insulin-like growth factors I and II (IGF-I and IGF-II) in the developing rat. Our studies span the period between early somite stages and full term. We have used oligodeoxyribonucleotide probes and obtained results which are both topographically precise and highly reproducible. The gene coding for IGF-II is predominant throughout development. It is strongly expressed in the liver and yolk sac. A variety of other tissues also expresses the IGF-II gene, especially many mesodermally derived structures in the process of differentiation. Many tissues do not express IGF genes. Thus no IGF mRNA was demonstrable in ectodermally derived structures, including the central and peripheral nervous systems as well as the skin and its derivatives.

Animals↗
PubMed1986

Location of gene expression in CNS using hybridization histochemistry.

In this chapter we have placed heavy emphasis on our own recent work to lay out a workable recipe for hybridization histochemistry. Only a trickle of papers followed the initial benchmark excursions into in situ labeling of tissue sections. Our own entry into this field was as late starters in 1978, but since then a confluence of important questions and technical advances has served to make hybridization histochemistry much more attractive and universally applicable as a research tool. Hybridization histochemistry allows the location of anatomical sites of gene expression and viral replication with unique specificity and is able to solve some problems for which there is no other suitable technique available in the central nervous system. For example, allowing that peptides may enter neurons by a variety of mechanisms and then be christened neuroendocrine peptides, it has become a compelling issue to know which cells are manufacturing the peptide. Thus, much can be learned by the approach elegantly demonstrated by Gee et al., of locating mRNA and its peptide product within the same neuron. The intracellular location of specific mRNA for a neuropeptide in a cell body indicates a very high probability that the peptide is secreted as a neurotransmitter or a neuromodulator from sites associated with the cell body. Our introduction of the use of whole mouse sections and large sections of brain of large animals in hybridization histochemistry has great potential in locating hormonal, enzymatic, and growth factor gene expression. The technique has been applied most elegantly by others to developmental studies and for the examination of viral infection. Resolution down to a single cell in heterogeneous tissue was beyond the original expectation of the capability of 32P-labeled probes and single cells in sections shown in Fig. 2 is probably the limit of resolution with this isotope. There is no reason why other isotopes, fluorescent labels, or labels suitable for EM should not take the resolution of the technique to intracellular. The horizon of application is widened enormously by the successful application of synthetic oligonucleotide probes, and at the same time unshackles the procedure from dependence upon a fully functional molecular biology laboratory. Although hybridization is a valuable research tool which we have applied to location of neuropeptides in the brain, it should soon find a niche in many fields and in a short time should become a key diagnostic tool.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1985

Hybridization histochemistry.

In this review we have used our own recent work as a flagship to illustrate the recent renaissance of interest in hybridization histochemistry. A trickle of papers followed the initial key excursion into the in situ labeling of tissue sections (48-50). Our own entry into this field started in 1978 and since then a confluence of important questions and technical advances has served to make hybridization histochemistry much more attractive as a research tool. Hybridization histochemistry is able to solve some problems for which there is no other suitable technique at this time. Hybridization histochemistry provides the location of anatomical sites of gene expression, and viral replication, with uniquely high specificity. We have taken 32P-labeled probes to what appears to be their limit of resolution, which is single cells in thin sections. While 32P has clear disadvantages, exposure time is relatively short and the use of fast-X-ray film to preview the results and estimate exposure time for emulsion has been turned to advantage. Our introduction (27) of the use of whole-mouse sections in hybridization histochemistry has great potential in hormonal, enzymatic, and growth factor gene expression and will no doubt prove of great use in developmental studies and examination of viral infection. The use of synthetic DNA (synthetic oligonucleotides) unshackles the technique from the need for an associated molecular biology laboratory and at once widens the horizon of application of the technique. Although hybridization histochemistry is a valuable research tool which will soon find a niche in many fields, in a short time it should become a key diagnostic aid. It may well become the method of preference for detection of the expression of oncogenes and other cancer-related genes and for viruses which for other reasons are difficult to detect.

Animals↗
PubMed1985

Renin gene expression in vessels of the ovine renal cortex.

Using hybridization histochemistry, a technique which localizes specific mRNA populations in tissue sections with a 700 base pair recombinant DNA probe which codes for ovine renin, we have localized renin gene expression in the afferent arteriole of the juxtaglomerular apparatus (JGA) in the sheep renal cortex. Specific labelling representing renin gene expression was also found at a distance from the glomerular tuft in the walls of the afferent arteriole and also in cells in the medial layer of larger vessels of the renal cortex, specifically the interlobular arteries. These observations provide morphological evidence of renin gene expression at these sites and, combined with ultrastructural and immunocytochemical evidence suggest that renin is synthesized and stored in the afferent arteriole either within the JGA or at a distance from the glomerulus, and in the smooth muscle coat of the interlobular arteries in the sheep kidney.

Animals↗
PubMed1984

Hybridization histochemistry: use of recombinant DNA for tissue localizations of specific mRNA populations.

A procedure of in situ hybridization, termed hybridization histochemistry has been developed to locate in specially prepared whole sections of tissue those areas which contain specific mRNA populations, the presence of which would indicate that a certain gene is "switched on" and production of a specific protein or peptide is highly probable. In the main 32P-labelled recombinant DNA probes were used. These probes were complementary to several peptide hormones or enzymes. 32P-labelled DNA probes were favoured initially because the short time needed for exposure enabled the development of the procedure to proceed quickly. The results indicate that the mRNA's of the species being examined are present in whole sections of tissue in a physical state which leaves them accessible to cDNA probes. The results are highly reproducible and specific. Because the method allows a clear distinction to be made between those cells which manufacture a peptide or protein and those which have imported the material it will have a special use in resolving many controversies over specific sites of origin of peptides and proteins, especially in tissues e.g. CNS and gut which are functionally complex and morphologically heterogeneous.

Animals↗
PubMedDate not supplied

Mouse prepro-epidermal growth factor synthesis by the kidney and other tissues.

Epidermal growth factor (EGF), a protein comprising 53 amino acids, is derived from a precursor of 1,217 amino acids that includes at least seven EGF-like sequences. EGF has diverse biological activities: it is a potent mitogen for many tissue culture cells, inhibits gastric acid secretion from the intestinal mucosa and promotes healing of the corneal epithelium. EGF given to fetal animals accelerates several developmental processes including palate formation, incisor eruption, eyelid opening and lung maturation. However, the physiological roles of EGF in vivo are unknown. The presence of high-affinity receptors in many fetal and adult tissues suggests that EGF is involved in normal cellular functions. Immunocytochemical studies have revealed the presence of EGF in mouse and human submaxillary glands, rat brain and human intestine. The low levels of EGF in extracts from many tissues may reflect sequestration rather than synthesis of the polypeptide. We show here that several mouse tissues contain preproEGF mRNA and that it is synthesized mainly in the distal tubules of the kidney. PreproEGF does not seem to be processed to EGF or other peptides in this tissue.

Animals↗
PubMedDate not supplied

Use of synthetic oligonucleotide and recombinant DNA probes to study renin gene expression.

Using hybridization histochemistry renin gene expression has been localized in the juxtaglomerular apparatus (JGA) of the renal cortex in both mouse and sheep kidney. This technique also located renin gene expression in afferent arterioles and interlobular arteries distant from the glomerular tuft in lamb renal cortex. A short (30 mer) synthetic oligonucleotide probe, complementary to a region of the mouse submaxillary gland renin gene, specifically labelled mouse submaxillary gland and kidney. Hybridization histochemistry and Northern blot analysis using both the synthetic oligonucleotide (mouse) probe and a 700 base pair recombinant (sheep) probe showed differences in renin gene expression in the kidney in response to Na restriction in the mouse and Na depletion in the sheep.

Animals↗
PubMedDate not supplied

Furosemide compounds enhance urinary excretion of active kallikrein independently of their effects on urinary electrolyte excretion.

The chemical and diuretic effects of furosemide on the excretion and activation of urinary prokallikrein were investigated in rats by treatment with furosemide compounds with a range of diuretic activity or amiloride hydrochloride or the vehicle. Diuresis occurred with furosemide and benzyl furosemide, but not with isofurosemide, amiloride hydrochloride, or the vehicle. The urinary excretion rate of active kallikrein was significantly elevated above controls throughout the 72 h of treatment with all of the drugs tested, regardless of the level of diuresis or the rate of urinary electrolyte excretion. In contrast, the urinary excretion rate of total kallikrein (prokallikrein + active kallikrein) was unchanged in all groups. These data indicate that furosemide derivatives increase the activation, but not the excretion, of urinary prokallikrein and that these effects are unrelated to the chemical structure or diuretic activity of the compounds or to overall changes in urinary electrolyte excretion rates.

Animals↗
PubMedDate not supplied

Furosemide treatment alters the distribution of kallikrein gene expression in kidneys of mice.

The renal kallikrein gene is normally expressed in convoluted distal and connecting tubules of the renal cortex. After furosemide treatment of mice there was strong expression also in the distal thick ascending limb of the loop of Henle in the renal outer medulla. This expansion of the kallikrein-expressing distal tubule cell population did not occur in mice on a sodium-restricted diet, in sodium-restricted mice given a high potassium diet or in untreated controls. Although the overall level of kallikrein gene expression was significantly greater in kidneys from furosemide-treated mice than in any other group, there was no detectable change in renal kallikrein gene expression in the submandibular glands after any of the treatments.

Animals↗