Keith A. Carson

Electron microscopic investigation of the effects of diabetes mellitus on the Achilles tendon

Fine structural changes in the Achilles tendons of patients with long-term diabetes mellitus were investigated. All patients had clinical and electrophysiological evidence of diabetic neuropathy and had ulceration and/or Charcot neuroarthropathy. Several differences between tendons of diabetic (n = 12) and nondiabetic (n = 5) individuals were observed by electron microscopy. In diabetics, these differences included increased packing density of collagen fibrils, decreases in fibrillar diameter, and abnormal fibril morphology. In one diabetic patient, individual collagen fibrils were tightly apposed so that many areas of tendon appeared as a single mass of closely adhering fibrillae. In addition, foci in which collagen fibrils appeared twisted, curved, overlapping and otherwise highly disorganized were common in specimens from most patients (11 of 12). These morphologic abnormalities in the Achilles tendons of diabetics appear to reflect a poorly known process of structural reorganization that may be the result of nonenzymatic glycation expressed over many years. Such structural changes could contribute to the tightening of the Achilles tendor a phenomenon consistent with clinical observations of extreme shortening of the Achilles tendon-gastrocnemius-soleus complex common in advanced diabetic neuropaths. In patients with diabetic neuropathy, tendon shortening causes severe equinus that may precipitate serious ulceration, stress fractures, and Charcot collapse of the foot. However, in nondiabetics, the fine structure of the Achilles tendon appears normal, consistent with the finding that the ultrastructural changes result from diabetes rather than neuropathy.

Electron microscopic localization of cholinesterase activity in Alzheimer brain tissue.

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity was localized by electron microscopic enzyme cytochemistry in cortex from Alzheimer brains and brains from non-demented cases. In the tangle-rich medial temporal cortex of the Alzheimer brain, most of the neuronal AChE was associated with neurofibrillary tangles. These structures also contained BChE activity. In normal neurons AChE activity was found in the rough endoplasmic reticulum, nuclear envelope and Golgi apparatus. Little BChE activity was noted in normal cortex. In neuritic plaques, AChE and BChE activity was associated mostly with the amyloid, but also with the neuritic component.

Ultrastructural evidence in mice that transganglionically transported horseradish peroxidase-wheat germ agglutinin conjugate reaches the intraspinal terminations of sensory neurons

Horseradish peroxidase conjugated to wheat germ agglutinin (HRP-WGA) has been localized by electron microscopy in the central projections of primary sensory neurons following injection of the tracer into the gastrocnemius muscle of mice. The tracer was taken up by nerve terminals in the muscle and transported transganglionically into the central spinal cord terminations of the primary sensory neurons. The HRP-WGA was localized using a new tetramethylbenzidine ultracytochemical method and the diaminobenzidine-glucose oxidase technique. The results demonstrate that the HRP-WGA is transported into the axon terminals after passing through the sensory ganglion. These data provide an experimental basis for the use of the conjugate to map the central projections of sensory neurons.

Chemically mediated behavior in Acari: Adapations for finding hosts and mates.

Ticks and mites respond to a limited spectrum of stimuli in their search for hosts and mates. Airborne chemical signals include carbon dioxide, ammonia, organic acids, terpenoids, 2,6-dichlorophenol, and other phenolic compounds. These are detected primarily by sensilla in and adjacent to Hallers organ. Most ixodid species examined have one or more multiporose sensilla that detect such volatiles. These olfactoreceptors enable the ticks to respond to remote volatile chemicals from hosts and from the other ticks, e.g., sex pheromones. Other sensilla, probably mechanogustatory in function, also occur on the tarsi. Gustatory sensilla on the palps detect assembly pheromones that enable ticks and mites to respond to conspecific or heterospecific chemical stimuli in their environment. Responses to those stimuli in ticks result in clustering, i.e., arrestant behavior. Arrestant behavior also occurs in certain mites. Finally, cheliceral chemosensilla enable ticks to recognize specific phagostimulants in host blood, e.g., ATP and glutathione, that stimulate feeding. InDennacentor variabilis andD. andersoni, these same cheliceral chemosensilla recognize species-specific genital sex pheromones in the vulvae of conspecific mates, without which they do not copulate.

Genital sex pheromones of ixodid ticks: 1. Evidence of occurrence in anterior reproductive tract of American dog tick,Dermacentor variabilis (Say) (Acari: Ixodidae)

The occurrence of a genital sex pheromone in the anterior reproductive tract of partially fed femaleDermacentor variabilis was demonstrated by extraction and bioassay. A new type of bioassay, the “neutered” female assay, was developed to test the potency of extracts or chemically defined fractions to stimulate males to copulate. Electrophysiological tests confirmed the ability of males to detect the pheromone with sensilla on their cheliceral digits. Males of bothD. variabilis andD. andersoni exhibited neuronal excitation when stimulated with extracts of theD. variabilis reproductive tissues. The pheromone, which is soluble in methanol, was fractionated and found to contain at least two fractions that stimulated copulation by sexually excited males. Evidently, the pheromone is a mixture of two or more compounds. Histologic, ultrastructural, and histochemical studies suggest the vestibular vagina as the site of genital sex pheromone occurrence, presumably from secretions of the surrounding lobular accessory gland. The identity of the compounds that comprise the pheromone remains unknown.

Seasonal Changes in Brown Fat and Pelage in Southern Short-Tailed Shrews

We examined cellular changes in interscapular brown adipose tissue and pelage characteristics in Blarina carolinensis collected throughout the year in eastern Virginia. Cellular volume occupied by mitochondria and maximum mitochondrial size were significantly greater in the brown adipose tissue of winter shrews than in summer shrews. Lipid droplets occupied greater volume and were larger in shrews in summer than winter shrews. There were no seasonal differences in hair density; Type I and Type II guard hairs were significantly longer in winter than summer by a factor of 1.3. Woolly hairs were 1.2 times longer in winter than summer, a non-significant difference.

Studies on possible role of catecholamines in regulation of sex pheromone gland activity in American dog tick,Dermacentor veriabilis (Say)

Administered monoamines affected sex pheromone activity in the foveal glands of the tick,Dermacentor variabilis (Say). Flooding the tissues of the female tick with reserpine, α-methyl-m-tyrosine methyl ester hydrochloride, and pilocarpine prior to feeding led to reductions in female sex attractant activity during engorgement. Similar treatments with cyclic AMP, dopamine, serotonin, 6-hydroxydopamine, and acetylcholine had no apparent effects on the attractiveness of feeding females. Assays (by gas chromatography) demonstrated substantial reductions in 2,6-dichlorophenol content following treatment with α-methyl-m-tyrosine methyl ester, pilocarpine, and, in most cases, with reserpine. Reserpine was effective only when administered in near-lethal concentrations to unfed females. In contrast, treatment with dopamine led to elevated 2,6-dichlorophenol content in most trials. X-ray microanalysis corroborated the evidence with reserpine and dopamine. These and other findings reported elsewhere implicate monoamines, presumably catecholamines, in the regulation of sex pheromone secretion in this species. The significance of these findings for understanding the physiological mechanisms involved in the regulation of sex pheromone secretion and biosynthesis is discussed.

Ultrastructural localization of acetylcholinesterase in the synganglion of the tick, Dermacentor variabilis (Say).

SummaryLight- and electron-microscopic enzyme cytochemistry was used to localize acetylcholinesterase (AChE) activity in the synganglion (brain) of the tick Dermacentor variabilis. High AChE activity was observed throughout the neuropil as well as adjacent to most neuronal perikarya. Intracellular activity was not observed by light microscopy. By electron microscopy, reaction product was localized at the plasma membrane of glia and neurons. Enzyme activity was not associated with the olfactory globuli neurons. In other types of neurons, small amounts of reaction product were observed in the Golgi apparatus and nuclear envelope. Large neurosecretory neurons contained activity that appeared to be associated with deep invaginations of the plasma membrane as well as intracellular membranes. AChE activity was also associated with processes of both neurons and glia. In most peripheral nerves AChE activity was associated with virtually all axons. Clearly then, AChE is associated with glia and non-cholinergic neurons as well as with presumed cholinergic neurons. The widespread localization and large amounts of AChE in the tick brain exceeds that reported for other invertebrates and vertebrates. As has been suggested for other animals, AChE in the tick brain may have functions in addition to its known role in cholinergic neurotransmission.