Jeffrey H. Spring

A Novel Target for Antidiuretic Hormone in Insects

Diuresis in insects is controlled by two antagonistic hormone groups: diuretic hormones, which promote water loss, and antidiuretic hormones, which inhibit it. All known antidiuretic factors act solely to promote fluid reabsorption by the hindgut and do not affect secretion by the Malpighian tubules. In the house cricket, Acheta domesticus, an antidiuretic hormone was found that inhibits fluid secretion by the Malpighian tubules but has no effect on the hindgut. Correlations were found between the density of neurosecretory granules and the presence of antidiuretic hormone in the corpora cardiaca, suggesting that the hormone is released from specific axons. Its release is triggered by dehydration; the hormone is detectable in the hemolymph of water-deprived crickets. These results imply that an unusual mechanism regulates water balance in these insects.

Excretion in the house cricket (Acheta domesticus): Fine structure of the Malpighian tubules

An ultrastructural analysis of the Malpighian tubules of the cricket, Acheta domesticus, is presented. The excretory system of the cricket is unusual in that the 112 Malpighian tubules do not attach directly to the gut, but fuse to form a bladder-like ampulla which is joined to the colon by a muscular ureter. The tubules have three structurally distinct segments and consist of four cell types. Attached by basal lamina to the outer surface of the distal tip are nodules, consisting of small cells on membranous stalks. These are presumed to serve as attachments to the body wall. The distal 20% of the tubule is hyaline, consisting of a monolayer of squamous cells that appear to be secretory. The mid-tubule comprises 75% of the total length and is the primary region for fluid secretion. It is also characterized by having large numbers of laminate spheres in the cytoplasm of the cells. The proximal 5% of each tubule consists of more columnar cells and may function in fluid resorption. The relationship between structural features and known physiological functions are discussed.

Ultrastructural changes in the Malpighian tubules of the house cricket, Acheta domesticus, at the onset of diuresis: A time study

The Malpighian tubules (Mt) of insects are responsible for maintaining osmotic homeostasis and eliminating waste from the hemolymph. When stimulated by diuretic factors the tubule cells are able to transport extraordinary volumes of fluid over short periods of time. We have been studying the changes that occur within the cells that accompany and facilitate this phenomenon. We present the ultrastructural changes that occur in the mid‐tubule of the house cricket, Acheta domesticus, following exposure to the second messenger analog, dibutyryl cAMP, over the period from 15–420 sec. Vacuolation of the cytoplasm begins as early as 30 sec poststimulation with a significant increase in vacuolation occurring after 120 sec. As expected, there is an increase in the surface area of the basolateral membrane to facilitate the rapid movement of fluid into the cells. Other ultrastructural changes noted to accompany the onset of diuresis include the movement of mitochondria into areas adjacent to transport membranes, the vesiculation of Golgi, mobilization of CaPO4 spherites, and a direct interaction of these spherites with active mitochondria. We discuss several possible roles for these changes in terms of rapid fluid transport. J. Morphol. 247:80–92, 2001.

Excretion in the house cricketAcheta domesticus: Effects of diuretics on the structure of the mid-tubule

Most structural studies of insect Malpighian tubules focus on freshly dissected tissue, which is in fact stimulated by diuretic factors. In this study, we examine tubules from the house cricket Acheta domesticus in four discrete secretory states: control (freshly dissected); unstimulated (held in vitro for 90 min prior to fixation); corpus cardiacum‐stimulated (held in vitro for 60 min, then stimulated with corpus cardiacum homogenates for 30 min prior to fixation); and cAMP‐stimulated (held in vitro for 60 min, then stimulated with dibutyryl cAMP for 30 min prior to fixation). In unstimulated tubules, we see a reduction in vacuolization and a near‐complete collapse of the basolateral infolds. Stimulated tubules show several major structural shifts: mitochondria are darkly stained with well‐defined cristae, there is extensive vacuolization of the tissue and expansion of the basolateral spaces, and the CaPO4 spherites appear to be ejected into the lumen. cAMP‐stimulated tubules showed the most pronounced structural changes, including the presence of a newly reported ultrastructural feature for A. domesticus Malpighian tubules, referred to here as paracrystalline arrays, which appear as stacks of membrane localized in the perinuclear region.

Membrane dynamics in the Malpighian tubules of the house cricket, Acheta domesticus

In Acheta domesticus, the Malpighian tubules (Mt) are composed of three morphologically distinct regions (proximal, mid and distal), each consisting of a single cell type. The bulk of the Mt is composed of the midtubule, which shows the greatest response to corticotropin releasing factor-related diuretic peptides (CRF-DP). We know from previous laboratory studies that the second messenger cAMP and its analog dibutyryl cAMP (db-cAMP) cause an approximate doubling in the secretion rate and that this is accompanied by notable ultrastructural changes in the midtubule, especially membrane reorganization in the basal area and extensive vesiculation of the cytoplasm. In this study, we examined the morphological changes in membranes both at the cell surface and internally. By enzymatically removing the basal lamina, we examined the increase in spacing between infolded membranes initiated by db-cAMP stimulation. To examine the intracellular membranes, we used a technique developed for use in invertebrate tissues. This allowed the removal of the cytoplasm for high resolution scanning electron microscopy (HR-SEM) while maintaining the integrity of the lipid constituents of the cell. By using HR-SEM and confocal laser scanning microscopy (CLSM), we gained a unique three-dimensional perspective of the complexity of the internal membrane system of the A. domesticus Mt in both the unstimulated and db-cAMP-stimulated states.

Excretion in the house cricket (Acheta domesticus): Ultrastructure of the ampulla and ureter

An ultrastructural analysis of the ampulla and ureter of the cricket, Acheta domesticus, is presented. The excretory system of the cricket is unusual in that the 112 Malpighian tubules do not attach directly to the gut, but fuse to form a bladder-like ampulla which is joined to the colon by a muscular ureter. The ampulla consists of two cell types, primary and regenerative. Primary cells secrete large numbers of membrane-bound vesicles into the lumen and also appear to be involved in fluid reabsorption. Regenerative cells are very small and form a layer just beneath the basal lamina of the ampulla. They are believed to differentiate and replace sloughed off primary cells. The ureter is a muscular tube lined with cuticle which connects the ampulla (endoderm) with the colon (ectoderm). The probable origin and significance of the morphological modifications of the excretory system are discussed.

Presence and Preliminary Characterisation of Factors Regulating Carbohydrate and Lipid Metabolism Isolated from the Corpus Cardiacum of the Eastern Lubber Grasshopper

There appears to be considerable conservation within the arthropods of the group of small neuropeptides which are known as the AKH/RPCH-family (Gade, 1986) because of their structural similarity to the locust adipokinetic hormone I (AKH I, Stone et al., 1976) and the crustacean red pigment-concentrating hormone (RPCH, Fernlund and Josefsson, 1972). The adipokinetic hormones (AKH I and AKH II) are stored in the corpus cardiacum (CC) of grasshoppers and their hyperlipaemic action is thought to be associated with the large increase in metabolic rate which accompanies sustained flight (e. g. Locusta, Schistocerca). We therefore decided to study the flightless and slow-moving lubber grasshopper, Romalea microptera, to determine whether it had the ability to synthesise neuropeptides of similar structure to AKH and, if so, what their intrinsic function might be.