The midline interlaminar ligament of the spine: An anatomical study.

INTRODUCTION: During routine cadaveric dissection, Simonds et al. in 2019 found a previously undocumented ligament, which they termed the midline interlaminar ligament (MIL), in 24 out of 36 (76.5%) lumbar spinal levels. The MIL is an unpaired ligament located between and distinctly separate from the right and left ligamenta flava (LF). The purpose of this study was to identify the presence or absence of the MIL in the cervical, thoracic, and lumbar spinal regions and obtain detailed measurements of the ligaments' toughness (R) and elastic modulus (E). MATERIALS AND METHODS: Intact preserved cadaveric vertebrae from C2 to the upper sacral region were dissected. Presence or absence of the MIL was documented, and length and width of each MIL were measured in situ. The R and E of the LFs from corresponding spinal segments were found for comparison. RESULTS: At least one MIL was observed in 90.3% (28) of specimens. Eighty-eight MIL's were observed out of 186 cervical intervertebral levels (0.5%), 371 thoracic intervertebral levels (5.9%), and 101 lumbar intervertebral levels (63.4%). The mean width and length of the MIL were 1.21 ± 0.36 and 16.37 ± 2.17 mm, respectively. The mean R of the MIL and the LF were 1390.27 and 2068.04 J m-2 , respectively. The mean E of the MILs and LFs was 46.78 ± 16.65 and 51.15 ± 21.68 MPa, respectively. CONCLUSIONS: Based on our findings, the MIL was present in the majority of vertebrae in our cadaveric population with a predominance for the lumbar region.

Examining persistence of acute environmental enrichment-induced anti-sucrose craving effects in rats.

A single, overnight (acute) environmental enrichment (EE; a large environment with conspecifics and novel objects) experience robustly decreases sucrose consumption (taking) and responsiveness to sucrose-paired cues (seeking) in rats. Persisting effects of acute EE on sucrose seeking and taking have not yet been identified. In the present study, rats were trained to self-administer a 10% sucrose solution paired with a compound tone + light stimulus for 10 days in 2-h sessions. We then examined the persistence of acute EE effects at reducing sucrose seeking and taking in a 12-h test immediately following acute EE (Exp. 1), or for 7 days with daily 1-h tests immediately following acute EE, or after a 24-h delay (Exp. 2). We also examined the persistence of acute EE effects on sucrose taking in rats responding on a PR schedule in 7 daily sessions following acute EE (Exp. 3). We found that acute EE was effective at reducing responding for both sucrose and a sucrose-paired cue, persisting throughout the 12-h test (Exp. 1). A reduction in sucrose seeking persisted for 24 h and a reduction in sucrose taking persisted for 72 h following acute EE plus a 24-h delay prior to testing (Exp. 2). Decreased PR responding for sucrose was observed following acute EE; this reduction persisted for 48 h (Exp. 3). These findings indicate that acute exposure to EE has persisting effects at reducing sucrose seeking and taking in rats. Acute EE may have translational value as a non-pharmacological intervention to curb sucrose craving.

Systemic injection of the DAD1 antagonist SCH 23390 reduces saccharin seeking in rats.

Conditioned cues can elicit drug- and sucrose-seeking behaviors that have been shown to depend on dopamine (DA) D1 receptors. If DAD1 receptors are also involved in seeking behavior in general, blocking these receptors should reduce seeking behavior for a non-caloric, non-drug of abuse reinforcer such as saccharin. Forty-six male Long-Evans rats lever pressed for 0.3% saccharin solution 1 h/day for 10 days. A lever response also activated a tone plus a white stimulus light. This compound stimulus lasted for 5 s. After 1 day of forced abstinence, rats received systemic (0, 1, or 10 µg/kg IP; n = 15-16 per group) injections of SCH 23390 15 min prior to extinction testing. Systemic SCH 23390 reduced saccharin seeking evidenced by a significant reduction in active lever responding and a significant reduction in the number of active lever-contingent deliveries of the tone + light cue following pretreatment with 10 µg/kg SCH 23390. The slope of responding across the Test session in this group was also significantly steeper, indicating that SCH 23390 may have reduced the persistence of saccharin seeking. The results indicate that DAD1 receptors are involved in saccharin seeking and generalize the previously demonstrated anti-seeking effects of DAD1 antagonism to a non-caloric, non-drug of abuse reinforcer.

Sucrose Abstinence and Environmental Enrichment Effects on Mesocorticolimbic DARPP32 in Rats.

Dopamine- and cAMP-regulated neuronal phosphoprotein 32 kDa (DARPP32) is a signaling molecule that could serve as a molecular switch, promoting or restraining sucrose seeking. We measured DARPP32 and pThr34 DARPP32 in the brains of male Long-Evans rats with a history of sucrose self-administration followed by 1 or 30 days of abstinence and exposure to either overnight (acute) or one month (chronic) environmental enrichment (EE). Brains were extracted following a 1 h cue reactivity test or no exposure to the test environment. Micropunches (prelimbic, infralimbic, and anterior cingulate areas of the medial prefrontal cortex, orbitofrontal cortex, dorsal striatum, nucleus accumbens, and ventral tegmental area) were then processed using Western blot. Abstinence increased, while EE decreased, sucrose seeking. DARPP32 and pThr34 DARPP32 levels were affected by testing, abstinence, and/or EE in most regions. Especially salient results were observed in the nucleus accumbens core, a region associated with relapse behaviors. Both acute and chronic EE reduced DARPP32 in the nucleus accumbens core and acute EE increased the ratio of phosphorylated to total DARPP32. Degree of DARPP32 phosphorylation negatively correlated with sucrose seeking. These findings demonstrate a potential role for DARPP32 in mediating the "anti-craving" effect of EE.

Effects of dopamine D1 and D2 receptor agonists on environmental enrichment attenuated sucrose cue reactivity in rats.

RATIONALE: Acute or chronic environmental enrichment (EE) reduces sucrose cue reactivity in rats. This effect may be mediated by dopamine receptors. OBJECTIVES: We examined whether dopamine D1 or D2 receptor agonism could reverse the EE effect. We also examined whether any reversal effects would vary with the incubation of sucrose craving. METHODS: Following 10 days (2 h/day) of sucrose self-administration, rats experienced either 1 or 30 days of forced abstinence and either overnight (acute) or 29 day (chronic) EE. D1 (SKF 81297; 0, 0.3, or 1 mg/kg) or D2 (quinpirole; 0, 0.1, or 0.3 mg/kg) agonist was administered systemically immediately prior to a subsequent 2-h cue reactivity test the next day (n = 9-12 per group). RESULTS: Dose-dependent effects were limited to the day 1 test. High doses of the agonists increased day 1 acute EE cue reactivity to levels comparable to control animals. On the day 30 test, SKF 81297 increased cue reactivity in acute EE, chronic EE, and control rats. In contrast, quinpirole resulted in similar cue reactivity for control and enriched rats, more from a reduction in responding by controls vs. a recovery of responding by EE-experienced rats. CONCLUSIONS: Both D1 and D2 receptors may be involved in the acute EE-mediated decrease in cue reactivity observed following 1 day of forced abstinence. In contrast, at 30 days of forced abstinence, D1 receptors may be critical in cue reactivity as SKF 81297 was effective at both restoring responding of enriched animals and potentiating responding of controls.

Effects of acute or chronic environmental enrichment on regional Fos protein expression following sucrose cue-reactivity testing in rats.

Exposure to environmental enrichment (EE) reduces sucrose seeking by rats with a history of sucrose self-administration. The present experiment examined whether acute or chronic EE also reduces brain Fos levels, a protein marker indicative of neuronal activation. Fos levels were also examined after either 1 or 30 days of forced abstinence to examine whether Fos levels vary with the incubation of sucrose craving. Fos expression was examined in 18 regions and was identified in brain slices using immunohistochemistry. Fos levels were higher in most regions after 30 days of forced abstinence and were decreased in most regions by either acute or chronic EE. Eleven regions had some statistically significant effect and/or interaction of EE or incubation on Fos; the most salient of these are listed here. In the prelimbic cortex, there was an incubation of Fos and EE reduced Fos at both forced abstinence time points. In contrast, in the orbitofrontal cortex, there was no Fos incubation but EE reduced Fos at both forced abstinence time points. An interaction of EE and incubation was observed in the anterior cingulate cortex and nucleus accumbens core and shell where Fos incubated but EE only decreased Fos at the day 30 forced abstinence time point. In contrast, in the dorsolateral striatum Fos incubated, but EE robustly decreased Fos expression at both forced abstinence time points. These differential expression patterns provide rationale for more detailed, site-specific molecular functional studies in how they relate to the ability of EE to reduce sucrose seeking.

Brief exposure to novel or enriched environments reduces sucrose cue-reactivity and consumption in rats after 1 or 30 days of forced abstinence from self-administration.

Environmental enrichment (EE) reduces drug and sucrose cue-reactivity in rats. In a previous study we reported that 1 month of EE (large cage, toys, and social cohorts) significantly reduced sucrose cue-reactivity. In the present study, we examined whether overnight (22 h) EE would be as effective. We also examined whether social enrichment (SE), enrichment alone (SoloEE), or exposure to an alternative environment (AEnv) might account for the EE effect. Rats self-administered 10% sucrose (.2 mL/delivery) in 10 daily 2-h sessions. Sucrose delivery was accompanied by a tone+light cue. Rats were then exposed to enrichment or alternative environment conditions overnight (acute) or for 29 days (chronic). Sucrose cue-reactivity was measured after this period of forced abstinence in a session identical to training, but no sucrose was delivered with the cue. All acute conditions markedly reduced sucrose cue-reactivity after 1 day of forced abstinence compared to single-housed rats in standard vivarium housing (CON). Sucrose consumption was also significantly reduced in all groups but SoloEE in a next-day test. All acute conditions but SE significantly reduced sucrose cue-reactivity when administered just prior to Day 30 of forced abstinence; all reduced sucrose consumption in a next-day test. All chronic conditions except for SE and AEnv significantly reduced sucrose cue-reactivity on the Day 30 test and sucrose consumption in a next day test. For both acute and chronic comparisons, EE manipulations were the most effective at reducing sucrose cue-reactivity and consumption. SoloEE and EE were equally effective at reducing sucrose cue-reactivity and similarly effective at reducing sucrose consumption. This indicates that social interaction is not a necessary condition for reducing sucrose-motivated behaviors. These results may be useful in the development of anti-relapse strategies for drug and food addictions.