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Painful neuropathy, like the other complications of diabetes, is a growing healthcare concern. Unfortunately, current treatments are of variable efficacy and do not target underlying pathogenic mechanisms, in part because these mechanisms are not well defined. Rat and mouse models of type 1 diabetes are frequently used to study diabetic neuropathy, with rats in particular being consistently reported to show allodynia and hyperalgesia. Models of type 2 diabetes are being used with increasing frequency, but the current literature on the progression of indices of neuropathic pain is variable and relatively few therapeutics have yet been developed in these models. While evidence for spontaneous pain in rodent models is sparse, measures of evoked mechanical, thermal and chemical pain can provide insight into the pathogenesis of the condition. The stocking and glove distribution of pain tantalizingly suggests that the generator site of neuropathic pain is found within the peripheral nervous system. However, emerging evidence demonstrates that amplification in the spinal cord, via spinal disinhibition and neuroinflammation, and also in the brain, via enhanced thalamic activity or decreased cortical inhibition, likely contribute to the pathogenesis of painful diabetic neuropathy. Several potential therapeutic strategies have emerged from preclinical studies, including prophylactic treatments that intervene against underlying mechanisms of disease, treatments that prevent gains of nociceptive function, treatments that suppress enhancements of nociceptive function, and treatments that impede normal nociceptive mechanisms. Ongoing challenges include unraveling the complexity of underlying pathogenic mechanisms, addressing the potential disconnect between the perceived location of pain and the actual pain generator and amplifier sites, and finding ways to identify which mechanisms operate in specific patients to allow rational and individualized choice of targeted therapies.

Abnormal neuronal calcium (Ca2+) homeostasis has been implicated in numerous diseases of the nervous system. The pathogenesis of two increasingly common disorders of the peripheral nervous system, namely neuropathic pain and diabetic polyneuropathy, has been associated with aberrant Ca2+ channel expression and function. Here we review the current state of knowledge regarding the role of Ca2+ dyshomeostasis and associated mitochondrial dysfunction in painful and diabetic neuropathies. The central impact of both alterations of Ca2+ signalling at the plasma membrane and also intracellular Ca2+ handling on sensory neurone function is discussed and related to abnormal endoplasmic reticulum performance. We also present new data highlighting sub-optimal axonal Ca2+ signalling in diabetic neuropathy and discuss the putative role for this abnormality in the induction of axonal degeneration in peripheral neuropathies. The accumulating evidence implicating Ca2+ dysregulation in both painful and degenerative neuropathies, along with recent advances in understanding of regional variations in Ca2+ channel and pump structures, makes modulation of neuronal Ca2+ handling an increasingly viable approach for therapeutic interventions against the painful and degenerative aspects of many peripheral neuropathies., (2009 Elsevier Ltd. All rights reserved.)

Since diabetes is a risk factor for Alzheimer's disease (AD), we asked if there is a functional interaction between high glucose and elevated beta amyloid peptide (Abeta) in cultured brain microvascular endothelial cells and presymptomatic AD transgenic mice. When cultured brain microvascular endothelial cells are exposed to both high glucose and low levels of Abeta, there is a synergistic interaction to cause an increased accumulation of advanced glycation products (AGE) and reactive oxygen species (ROS). When presymptomatic mice expressing mutant human amyloid precursor protein and presenilin are made diabetic, they have a decrease in cognitive function relative to control mice. Associated with the cognitive deficit are increases in brain microvascular AGE and iNOS expression, and the loss of the synaptic spine protein drebrin. No amyloid plaques or tangles are observed within the brains of any group. These data show that diabetes causes a synergistic potentiation of some indices of AD in transgenic animals that are presymptomatic for the classical features of the disease.

The effects of hindlimb temperature on sciatic nerve and skeletal muscle laser Doppler vascular conductance (LDVC) were assessed in anesthetized control and streptozotocin (STZ)-diabetic rats. With core temperature at 37 degrees C and exposed hindlimb temperature at 32 degrees C, nerve LDVC was significantly lower in rats after 8 weeks of STZ diabetes than in age-matched control rats. Subsequent warming of the exposed hindlimb of control rats from 32 degrees C to 37 degrees C significantly decreased nerve LDVC by 41% and increased muscle LDVC by 48%. Because nerve LDVC was unchanged by hindlimb warming in STZ-diabetic rats, there was no significant difference between control and diabetic nerve LDVC at 37 degrees C. In a second study, after 6 weeks of STZ diabetes, changes from control nerve LDVC were shown to depend on temperature rather than the duration of surgical exposure. These findings emphasize that information about hindlimb temperature is a prerequisite for interpreting the effects of experimental diabetes on hindlimb nerve blood flow.

The association between mast cells and vessel morphometry in sciatic nerve was examined after seven days in animals fed a diet of 40% D-galactose and compared to control rats and to galactose-fed animals treated with the aldose reductase inhibitor, Tolrestat. Electron microscopy revealed an increase in the total number of mast cells and the number of degranulated mast cells in galactose-fed animals (7.8 +/- 2.9; 2.6 +/- 2.9; mean +/- SD) compared to controls (4.6 +/- 2.1; degranulated mast cells were not seen in any control nerves) and Tolrestat-treated, galactose-fed animals (4.4 +/- 2.5; 0.1 +/- 0.4). Although no significant differences were noted in the numbers of vessels between the three groups, an index of vasoconstriction was significantly increased in the galactose-fed animals (0.115 +/- 0.048; mean +/- SD) compared to controls (0.068 +/- 0.011) and Tolrestat-treated, galactose-fed animals (0.075 +/- 0.20). These data suggest that mast cell degranulation is associated with the vascular constriction induced by seven days of galactose intoxication and that both may be prevented by inhibiting aldose reductase.

This study was designed to assess the effects of experimental diabetes and nerve crush injury upon vasoactive intestinal polypeptide (VIP) content and axonal transport in the dorsal root ganglia (DRG) and sciatic nerve. Sciatic nerve crush injury in control and 3-week streptozotocin-diabetic rats was followed 6.5 days later by placement of 2 constricting ligatures above the site of injury. After 12 h, the L4 and L5 DRG and sciatic nerve were removed for VIP radioimmunoassay. Similar samples were also taken from control and diabetic rats whose nerve had been ligated without a preceding crush. VIP was increased over 2-fold in ganglia and 4-fold in nerves of crush-injured controls compared to uninjured controls (both P < 0.01). Crush injury also increased ganglion and nerve VIP in diabetic rats (P < 0.05 and 0.01, respectively) but the increase was less than what occurred in crush-injured controls (both P < 0.05). The accumulation of VIP proximal to a sciatic ligature was similar in control and diabetic rats and was not altered in either group by crush injury, while retrograde transport of VIP was initiated by crush injury in both control and diabetic rats. These data show that short-term diabetes does not alter the amount and peripheral axonal transport of VIP in the sciatic nerve but impairs the ability of peripheral nerve to respond to injury.

The role of the enzyme aldose reductase in nerve homeostasis was examined by treating rats with an aldose reductase inhibitor. Female Sprague-Dawley rats were treated with Ponalrestat (25 mg/kg/day) or with excipient alone for 4 to 12 weeks before examining electrophysiologic function, endoneurial fluid electrolyte concentrations, nerve polyol levels, water content and (Na+,K+)-ATPase activity. Sorbitol, the product of glucose metabolism by aldose reductase, was detected in all nerves from control animals, whereas it was below detection limits in 7 of 11 nerves from Ponalrestat-treated rats. Ponalrestat treatment reduced endoneurial fluid sodium and chloride concentrations by 25% and 37%, respectively (both P < 0.001). No differences in nerve water content, conduction velocity, or ATPase activities were detected. These data, and previous studies demonstrating that increased flux through aldose reductase causes the accumulation of endoneurial electrolytes, suggest a role for this enzyme in modulation of the endoneurial microenvironment. However, short-term inhibition of aldose reductase does not appear to affect nerve function. Thus, our findings do not elicit concerns regarding the use of aldose reductase inhibitors in the treatment of clinical diabetic neuropathy.

This study describes the electrophysiological responses of endoneurial preparations derived from rat sciatic nerve to acute hypoxia in vitro. Preparations from control rats exhibited a 40% decline in compound action potential (CAP) amplitude after 40 min exposure to medium gassed with 8% O2. In preparations from 4 week streptozotocin-diabetic rats CAP declined by only 29%, indicating a resistance to hypoxic conduction blockade. Treating diabetic rats with mixed bovine brain gangliosides (10 mg/kg/day i.p.) exaggerated this resistance to hypoxic conduction blockade as CAP amplitude fell to only 18% of initial values. In a separate experiment, treating non-diabetic rats with gangliosides (10 mg/kg/day i.p.) or adding gangliosides (400 micrograms/ml) directly to the medium in which control nerves were maintained during in vitro recording also significantly attenuated the decline in CAP amplitude after 40 min hypoxia, thus effectively inducing a resistance to hypoxic conduction blockade similar to that observed in nerves from diabetic rats. These studies demonstrate that the systemic or acute local administration of gangliosides induces a resistance to hypoxic conduction block in normal nerve and exaggerates the resistance to hypoxic conduction block of diabetic rats.

This study examined sciatic motor nerve conduction velocity (MNCV) and the accumulation of 6-phosphofructokinase (PFK) activity proximal and distal to a sciatic nerve ligature in rats fed a diet containing 20% galactose for 4 weeks. MNCV was reduced in the galactose-fed rats to 94% of controls, (P less than 0.05) but PFK activity accumulations were not different from those in controls. Daily administration of the aldose reductase inhibitor ponalrestat throughout the study to another group of galactose-fed rats prevented dulcitol accumulation, myo-inositol depletion and increased water content of the sciatic nerve seen in galactose-fed rats. This effective aldose reductase inhibition also prevented the reduced MNCV and had no effect on accumulations of PFK activity. These data support earlier work suggesting that deficits in the axonal transport of PFK activity in diabetic rats are unrelated to either exaggerated flux through the polyol pathway, polyol accumulation or ischemic hypoxia and indicate the possible need for the elucidation of other pathogenic mechanisms which may contribute to the development of diabetic neuropathy.

This study describes the electrophysiological responses of endoneurial preparations derived from rat sciatic nerve to acute hypoxia in vitro. Preparations from control rats exhibited a marked decline in compound action potential (CAP) amplitude coupled with an increase in latency, during 40 minutes exposure to 8% O2. In contrast, preparations from 4 week streptozotocin-diabetic rats showed a greatly reduced decline in CAP amplitude, with an increase in latency. Twice-daily insulin treatment of diabetic rats resulted in a pattern of CAP amplitude decline that initially resembled that of untreated diabetics but by 40 min was similar to controls, with latency again increasing during hypoxia. Nerves were also maintained in 25 mmol/l glucose, rather than the 5 mmol/l glucose of the above studies. Under such conditions the performance of nerves from diabetic rats was unaltered. Nerves from control rats exhibited an initial resistance to hypoxia but by 40 min CAP had declined to values of control rats maintained in 5 mmol/l glucose. An increase in latency during hypoxia was also noted in preparations from control or diabetic rats maintained in 25 mmol/l glucose. The maintenance of CAP amplitude during hypoxia by diabetic preparations is initially related to increased substrate availability, with an additional component that is not related to external glucose levels in vitro, and is absent after insulin treatment of diabetic rats.

This study examined the effect of treatment of control and streptozotocin-diabetic rats with a mixture of gangliosides, derived from bovine brain, on parameters of axonal transport of substance P-like immunoreactivity (SPLI) and its levels in sciatic nerve and lumbar spinal ganglia. Rats were treated daily (10 mg/kg i.p.) for 28 days and compared with untreated control and diabetic groups. The duration of diabetes was 28 days in both cases. Untreated diabetic rats showed deficits in accumulation of axonally transported SPLI proximal (59% of controls) and distal (34% of controls) to sciatic nerve ligations (left in place for 12 h). Rates of accumulation were unaltered by diabetes. There were small numerical reductions in the SPLI content of unconstricted sciatic nerve and of L4 and L5 dorsal root ganglia in diabetic rats. None of these diabetes-associated changes was altered by ganglioside treatment, nor was there any indication of an effect of gangliosides on substance P in non-diabetic rats. The implications are discussed in relation to the possible pathogenesis of diabetic neuropathy.

This study measured the content of substance P-like immunoreactivity (SPLI) in peripheral nervous tissue (lumbar dorsal root ganglia, sciatic nerve), skin (snout, foot), gastrointestinal tract (stomach, terminal ileum) and in the atria of the heart. Animals studied were long-term (11 months) streptozotocin-diabetic rats compared with age-matched control rats. All diabetic rats were given a very long acting insulin preparation twice weekly to reduce morbidity. Half of the diabetic rats were given the aldose reductase inhibitor, sorbinil (mean dose 30 mg/kg/day body weight by dietary admixture) over the entire protocol. Diabetic rats (given insulin only) showed marked accumulation of sorbitol and fructose together with myo-inositol depletion in their sciatic nerves. The sciatic nerves of the sorbinil-treated diabetic rats contained amounts of sorbitol, fructose and myo-inositol which were similar to those of non-diabetic rats, in spite of large amounts of nerve glucose in the sorbinil-treated animals. Thus, the inhibition of aldose reductase was successful. The L4 and L5 dorsal root ganglia of the diabetic rats showed reduced SPLI (63% and 72% respectively of control ganglia; P less than 0.05). There was also numerical reduction in sciatic nerve SPLI (84% of control nerve). There were no effects of sorbinil treatment on the reduced SPLI levels in ganglia or sciatic nerve. In the gastrointestinal tract the levels of SPLI were reduced in diabetic rats even when data were adjusted to take account of tissue hypertrophy (diabetic SPLI/whole stomach was 60% controls, P less than 0.01 and SPLI/cm ileum was 78%, though the latter did not attain statistical significance). In skin SPLI/unit area was raised in the diabetic rats to 145% of controls for foot skin and 151% for snout skin. Changes in SPLI content of gastrointestinal tract were unaffected by sorbinil treatment; in the skin the elevations were enhanced to 188% and 270% of respective control values for foot and snout skin. The SPLI content of the atria was unaffected by diabetes or sorbinil. These data are not consistent with a generalised impairment of delivery of substance P by axonal transport in experimental diabetes; special factors appear to influence the levels in neurones innervating different tissues. Exaggerated flux through the polyol pathway appears to be uninvolved.

Ouabain-sensitive ATPase activity (expressed as nmol ADP produced/h/mg (wet) nerve +/- SEM) was measured in homogenates of sciatic nerve from control rats and rats with streptozotocin-induced diabetes of 8 wk duration. Nerves from diabetic rats showed activity (21.7 +/- 2.0) which was significantly (p less than 0.05) less than that of controls (34.6 +/- 4.8). These animals also showed a deficit in conduction velocity (m/sec +/- SEM) of sciatic nerve motoneurones (50.7 +/- 0.4 vs. 57.7 +/- 0.7 in controls; p less than 0.001). In parallel, matched control and diabetic groups were treated daily with mixed gangliosides extracted from bovine brain (10 mg/kg i.p.). After such treatment for 8 wk the deficit in ouabain-sensitive ATPase activity did not develop in the diabetic group (treated diabetics, 31.9 +/- 3.7; treated controls, 34.5 +/- 3.8). However, the treatment did not affect the deficit in motor nerve conduction velocity (treated diabetics, 50.9 +/- 1.1 vs. treated controls, 57.9 +/- 0.5; p less than 0.001). Accumulations of the polyol pathway metabolites--sorbitol and fructose--together with depletion of nerve myo-inositol were similar in both diabetic groups. These data indicate an etiology for the conduction velocity deficit which differs from that of the deficit in ouabain-sensitive ATPase.