Elevated granulocyte colony stimulating factor (CSF) causes cerebellar deficits and anxiety in a model of CSF-1 receptor related leukodystrophy.

Colony stimulating factor (CSF) receptor-1 (CSF-1R)-related leukoencephalopathy (CRL) is an adult-onset, demyelinating and neurodegenerative disease caused by autosomal dominant mutations in CSF1R, modeled by the Csf1r+/- mouse. The expression of Csf2, encoding granulocyte-macrophage CSF (GM-CSF) and of Csf3, encoding granulocyte CSF (G-CSF), are elevated in both mouse and human CRL brains. While monoallelic targeting of Csf2 has been shown to attenuate many behavioral and histological deficits of Csf1r+/- mice, including cognitive dysfunction and demyelination, the contribution of Csf3 has not been explored. In the present study, we investigate the behavioral, electrophysiological and histopathological phenotypes of Csf1r+/- mice following monoallelic targeting of Csf3. We show that Csf3 heterozygosity normalized the Csf3 levels in Csf1r+/- mouse brains and ameliorated anxiety-like behavior, motor coordination and social interaction deficits, but not the cognitive impairment of Csf1r+/- mice. Csf3 heterozygosity failed to prevent callosal demyelination. However, consistent with its effects on behavior, Csf3 heterozygosity normalized microglial morphology in the cerebellum and in the ventral, but not in the dorsal hippocampus. Csf1r+/- mice exhibited altered firing activity in the deep cerebellar nuclei (DCN) associated with increased engulfment of glutamatergic synapses by DCN microglia and increased deposition of the complement factor C1q on glutamatergic synapses. These phenotypes were significantly ameliorated by monoallelic deletion of Csf3. Our current and earlier findings indicate that G-CSF and GM-CSF play largely non-overlapping roles in CRL-like disease development in Csf1r+/- mice.

Scleral Tonometry Precision During Scleral Lens Wear: A Pilot Study.

PURPOSE: To evaluate the reproducibility, and therefore the utility, of using traditional tonometry devices for measuring intraocular pressure (IOP), while a prosthetic replacement of the ocular surface ecosystem device (PD) or scleral lens is applied to the eye. PATIENTS AND METHODS: Twenty subjects (40 eyes) with keratoconus were enrolled. With PD applied, the first 10 consecutive patients had IOP measured multiple times with a handheld tonometer (Tono-Pen AVIA, Reichert, Depew, NY) on the superotemporal sclera 1 mm posterior to the PD edge. This identical procedure was repeated for the next 10 consecutive patients with a pneumatonometer (Model 30, Reichert, Depew, NY). Once three reliable measurements, as defined by the study protocol, were obtained for an eye, the procedure was repeated with the same tonometer device on the fellow eye. RESULTS: The mean standard deviation for reliable IOP measurements was ±2.92 mmHg, median (IQR) of 2.62 (1.68 to 3.53) mmHg in the handheld tonometer group and ±1.98 mmHg in the pneumatonometer group. There was no statistically significant difference between the groups (p = 0.07). The mean IOP range for the reliable IOP measurements was 5.5 ± 3.80 mmHg, median (IQR) of 5 (3 to 7) mmHg for the handheld tonometer group and 3.71 ±1.12 mmHg in the pneumatonometer group. There was no statistically significant difference between the groups (p = 0.06). CONCLUSION: Handheld tonometry and pneumatonometry have poor reproducibility when used to measure scleral IOP in keratoconus patients, while a PD is applied to the eye. An alternative research model and methodology should be investigated and confirmed to have precision prior to proceeding with further analysis of any relationship between scleral lens wear and IOP.

Microglial reduction of colony stimulating factor-1 receptor expression is sufficient to confer adult onset leukodystrophy.

Adult onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a dementia resulting from dominantly inherited CSF1R inactivating mutations. The Csf1r+/- mouse mimics ALSP symptoms and pathology. Csf1r is mainly expressed in microglia, but also in cortical layer V neurons that are gradually lost in Csf1r+/- mice with age. We therefore examined whether microglial or neuronal Csf1r loss caused neurodegeneration in Csf1r+/- mice. The behavioral deficits, pathologies and elevation of Csf2 expression contributing to disease, previously described in the Csf1r+/- ALSP mouse, were reproduced by microglial deletion (MCsf1rhet mice), but not by neural deletion. Furthermore, increased Csf2 expression by callosal astrocytes, oligodendrocytes, and microglia was observed in Csf1r+/- mice and, in MCsf1rhet mice, the densities of these three cell types were increased in supraventricular patches displaying activated microglia, an early site of disease pathology. These data confirm that ALSP is a primary microgliopathy and inform future therapeutic and experimental approaches.

Microglial Homeostasis Requires Balanced CSF-1/CSF-2 Receptor Signaling.

CSF-1R haploinsufficiency causes adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). Previous studies in the Csf1r+/- mouse model of ALSP hypothesized a central role of elevated cerebral Csf2 expression. Here, we show that monoallelic deletion of Csf2 rescues most behavioral deficits and histopathological changes in Csf1r+/- mice by preventing microgliosis and eliminating most microglial transcriptomic alterations, including those indicative of oxidative stress and demyelination. We also show elevation of Csf2 transcripts and of several CSF-2 downstream targets in the brains of ALSP patients, demonstrating that the mechanisms identified in the mouse model are functional in humans. Our data provide insights into the mechanisms underlying ALSP. Because increased CSF2 levels and decreased microglial Csf1r expression have also been reported in Alzheimer's disease and multiple sclerosis, we suggest that the unbalanced CSF-1R/CSF-2 signaling we describe in the present study may contribute to the pathogenesis of other neurodegenerative conditions.